中华绒螯蟹的基因表达谱及生殖相关基因的克隆、表达模式研究
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摘要
中华绒螯蟹(Eriocheir sinensis)是中国淡水渔业的重要组成部分,它已经成为中国的一道传统美食。由于它肉质细嫩,味道鲜美,营养丰富,在全球市场供不应求且价格不菲。它是我国重要的经济水产养殖品种,该蟹的养殖已成为我国水产养殖的支柱产业。在过去的几十年中,随着密集化养殖的发展,由细菌、病毒和类立克次氏体属微生物所引发的各种疾病,以及性早熟、个体小型化以、种质退化等现象在中华绒螯蟹种群中频频发生,并且对螃蟹的质量造成灾难性损失,已经成为限制该蟹养殖业发展的重要瓶颈。因此认为了解其免疫防御机制、免疫因子特征,以及生殖生物学尤其是生殖的分子机理的研究,对螃蟹养殖的疾病控制和健康管理很有帮助,并为该蟹养殖过程中的人工生殖调控提供理论保障。本论文试图从分子角度来研究中华绒螯蟹的免疫防御机制以及雄性生殖机理,以期能有效突破中华绒螯蟹人工养殖的瓶颈,进一步促进该蟹养殖业的持续发展。
本研究借助分子生物学和生物信息学手段,首次完成对中华绒螯蟹肝胰腺cDNA文库的构建和表达序列标签(EST)的分析,并利用先进的Solexa测序技术,对中华绒螯蟹副性腺组织进行转录组及microRNA的大规模测序并分析其基因表达情况。不仅丰富了中华绒螯蟹的基因组数据库,同时筛选出许多免疫相关及生殖相关的的基因序列。通过对所测高通量数据分析的注释结果,结合已有物种生殖相关基因的研究现状,分析、筛选与该蟹生殖发育相关的部分重要基因；利用RACE和WORKING等技术,获得这些基因的cDNA全长以及基因的结构；利用realtime PCR研究不同基因在不同组织中以及同一组织不同发育阶段的表达情况,探讨这些基因在整个生殖发育过程中的调控作用。这些研究为了解中华绒螯蟹生殖机制提供了基础资料,同时也为深入探讨该蟹雄性生殖机理提供了重要的参考。主要研究结果如下：
1采用常规cDNA文库构建技术,成功构建了一个中华绒螯蟹肝胰腺的EST文库并且获得了3,279高质量序列,代表着1177个一致性序列。超过一半的一致性序列被鉴定是这一物种的未知的新基因。有很大一部分一致性序列(49%)在新发布的数据库中找到它们的同源物,这些同源物对于进一步的功能研究很重要。总结了与代谢和免疫相关的基因,借以考察肝胰腺的两种重要功能：代谢和免疫。为中华绒螯蟹的深入研究打下基础,包括转录组、物理图谱和整个基因组测序。展示了中华绒螯蟹与免疫密切相关的转录组和基因的重要性质。本文重点找到了与机体免疫相关的基因,分为5大类,包括蛋白酶类、抗菌肽、压力蛋白、抗氧化蛋白和其它与免疫相关的基因进行重点分析。蛋白酶类和他们的抑制剂在无脊椎动物抗感染反应中的重要性越来越明显。它们可能在应对侵染性蛋白的体液免役反应过程中表达,这些侵染性蛋白是由侵染性抗原产生的。文库中发现了1种抗菌肽,抗脂多糖因子。压力蛋白
包括铁蛋白、金属硫蛋白、热激蛋白70、胞溶质蛋白A。硒与谷胱甘肽系统在抗氧化防御体系。以及p-1,3-葡聚糖结合蛋白、C型凝集素、血蓝蛋白、干扰素等,都对中华绒螯蟹的免疫有着重要作用。
2从雄性中华绒螯蟹的副性腺组织中提取并分离mRNA,经超声破碎和随机引物反转成cDNA片段,利用Solexa测序(sequencing by synthesis)技术,对这些组织的RNA转录组进行全面测序；用生物信息学的手段和方法,对测序数据进行转录谱分析,分析各组织的基因表达情况,进一步综合各组织之间基因表达的相关性,探讨不同组织在该蟹雄性生殖过程中的作用及调控模式。共获得33,221,284条reads,包含2,657,702,720nt碱基,GC含量55.19%。通过软件组装拼接得到85,913条contig,将所有contig按序列长度从小到大排序,其中40,977条(47.70%) contig集中在100-200nt的长度.从第一条序列开始累加计算contig总长,所有contig的总长度为18,750,722。通过对Unigene的功能注释,其中有1713条基因条注释到General function prediction only,注释上Translation, ribosomal structure and biogenesis有952条基因,与Transcription相关的有869条Unigene。关于Posttranslational modification, protein turnover, chaperones和Replication, recombination and repair的基因分别有802和789条。KEGG是基因组破译方面的数据库。附性腺有1704条基因(14.99%)参与Metabolic pathways,675条基因参与Regulation of actin cytoskeleton。Spliceosome有564(4.96%)条基因参与其中。得到原始小RNA的fq数据后,对其进行去接头,去低质量reads,去污染等处理,得到干净的序列7939380条。统计小RNA(sRNA)的种类(用unique表示)及数量(用total表示),并对小RNA做长度分布统计,一般来说,小RNA的长度区间为18-30nt,通过长度分布的峰我们可以判断小RNA的种类,miRNA集中在21或22nt, siRNA集中在24nt, piRNA集中在30nt。共有Total sRNAs 7939380条,其中miRNA有382650条(4.82%)、rRNA有214013条(2.70%)、snRNA有2573条(0.03%)、snoRNA有563条(0.01%)、rRNA有432171条(5.44%)以及unann有6907410条(87.00%)。共有Unique sRNAs 2268484条,其中miRNA有23021条(1.01%)、rRNA有33764条(1.49%)、snRNA有1003条(0.04%)、snoRNA有224条(0.01%)、tRNA有29425条(1.30%)和unann有2181047条(96.15%)。
3生殖相关蛋白及其功能的研究是当前甲壳动物生殖生物学的研究重点之一,许多生殖相关蛋白及其功能已经明确。瘦素受体在生殖发育过程有调节生殖发育的作用。本研究根据EST提供的信息,采用RACE技术成功克隆了中华绒螯蟹瘦素受体蛋白(EsLEPR)cDNA全长。生物信息学分析发现,该基因cDNA全长序列的长度大小1403 bp包括一个72 bp的5'非翻译区(5' UTR),1929 bp的3'-非翻译区(3'UTR)的一个AATAAA加尾信号和23 bp的polyA尾,和402 bp的开放阅读框(ORF),编码133个氨基酸的多肽与预测分子量14.42kDa和理论等电点为4.46。其基因组DNA序列为3553bp其中有3个内含子和4个外显子组成,包含有典型的(GT和AG)二核苷酸内含子交界处的结构序列剪接位点。氨基酸序列中有一个保守区域：Vacuolar protein sorting 55 (Vps55)。133个氨基酸的序列中的保守区域含有4个跨膜束片段,分别在第4到第26个氨基酸,第33到第52,第67到第89,和第101到第123。氨基酸序列的多重比对结果显示与Eriocheir sinensis同源性最高的是(64%)丽蝇蛹集金小蜂(Nasonia vitripennis)。它与非洲爪蛙(Xenopus laevis,63%)、红鳍东方触(Takifugu rubripes,58%)、人(Homo sapiens,57%)、牛(Bos taurus,57%)、(Rhodnius prolixu,56%)、褐鼠(Rattus norvegicus,55%)、海鞘(Ciona intestinalis,54%),、猕猴(Macaca mulatta,54%),、蚜虫(Acyrthosiphon pisum,48%)和(Boltenia villosa,40%)都有较高的同源性。采用实时定量RT-PCR的方法分析中华绒螯蟹瘦素受体蛋白在不同组织中以(3-actin作为内参的相对表达情况。组织中的表达分析显示,中华绒螯蟹瘦素受体蛋白的mRNA在各个组织中均有表达,主要是在肠、精巢、胸神经节、脑神经节和肝胰腺等组织中表达。
4蜕皮激素引起的幼体蜕皮发育有关的各种活动。蜕皮激素调节蛋白可能是甲壳动物变态蜕皮级联反应的重要元素。在中华绒螯蟹养殖中性早熟和蜕皮未遂的问题经常发生。这两种病害与甲壳动物的蜕皮是密切相关的,研究蜕皮相关基因对于解决中华绒螯蟹性早熟和蜕皮未遂的问题有重要的意义。通过EST分析得到蜕皮激素调节蛋白的两条ESTs (FG358768,FG359773)；用RACE方法得到中华绒螯蟹蜕皮激素调节蛋白cDNA全长。中华绒螯蟹蜕皮激素调节蛋白基因cDNA全长序列的长度大小1384 bp包括一个117-bp的5'非翻译区(5'UTR)、841-bp的3'-非翻译区(3'UTR)的一个AATAAA加尾信号和31-bp的polyA尾,和426 bp的开放阅读框(ORF),编码141个氨基酸的多肽与预测分子量15,150.3 Da和理论等电点为3.86。中华绒螯蟹蜕皮激素调节蛋白的基因组DNA序列为3060bp其中有1个内含子和2个外显子组成。中华绒螯蟹蜕皮激素受体基因组序列的包含有典型的(GT和AG)二核苷酸内含子交界处的结构序列剪接位点。而且在基因组序列序列与其cDNA序列一致。用ClustalW软件对不同物种的蜕皮激素调节蛋白的氨基酸序列进行比对。中华绒螯蟹的蜕皮激素调节蛋白与Litopenaeus vannamei的蜕皮激素调节蛋白(ABD65303)同源性为34.4%,与Bombyx mori蜕皮激素调节蛋白(NP_001134647)的同源性为25.8%。序列比对结果显示有6个高度保守的半胱氨酸残基(Cys22, Cys36, Cys41, Cys87, Cys92, Cys133)。SMART程序分析表明,推导的氨基酸序列包含一个保守的结构域:the Niemann-Pick type C2(属于ML家族)结构域。是由119个氨基酸组成开始在Tyr19并在Val138结束。在ML保守结构域上有22个公认的胆固醇/脂质结合位点。采用SWISS-MODEL模型预测中华绒螯蟹蜕皮激素调节蛋白的三级结构,根据与其他物种相同蛋白很高的相似性,建立的三级结构的模板为:lnepA。模拟结构的氨基酸残基范围开始于第16个残疾和结束于第141残基和序列结构同源性为28.9%。在整个中华绒螯蟹蜕皮激素调节蛋白推导的氨基酸序列结构主要是β-折叠。采用实时定量RT-PCR的方法分析中华绒螯蟹蜕皮激素调节蛋白在不同组织中以β-actin作为内参的相对表达情况。组织中的表达分析显示,中华绒螯蟹蜕皮激素调节蛋白的mRNA主要是在肝胰腺组织中表达,在脑神经节中表达相对较高,在性腺、胃、肌肉和胸神经节中表达较少,而在心,鳃,血淋巴和肠中不表达。在肝胰腺中的表达最丰富,这与肝胰腺的功能密切相关,肝胰腺是蜕皮现象的主要指示器官。采用实时定量RT-PCR的方法,分析中华绒螯蟹蜕皮激素调节蛋白在中华绒螯蟹不同蜕皮时期(蜕皮前、蜕皮中、蜕皮后和硬壳期)肝胰腺中的表达情况。中华绒螯蟹蜕皮激素调节蛋白在蜕皮前的肝胰腺中表达量最高,在进入蜕皮时显著降低,蜕皮后开始回升,直到硬皮期回升到正常水平。
The mitten crab (Eriocheir sinensis) (Henri Milne Edwards 1854), which belongs to Crustacea, Decapoda, Grapsidae, Eriocheir, is a unique species of crab that is native to China. This species is a traditional savory food and therefore comprises one of the economical aquatic species of China. The farming of E. sinensis has been developing rapidly in China over the last two decades and reached its highest yield of 4.0 x 105 tonnes in 2005. With the development of intensive culture, various diseases caused by bacteria, viruses, or rickettsia-like organisms and the sexual precocity, individual small, germplasm degradation are frequent in cultured species populations and have caused catastrophic losses. It has become a major bottleneck in the development of aquaculture industry. Understanding the immune defence mechanisms of the species; characterizing of immune effectors, and the study of reproductive biology, especially the molecular mechanism of reproductive are believed to be helpful in health management and disease control in crab aquaculture, and provide a theoretical to the reproductive control of the artificial crab breeding process.
In the present study, we presented the methodology and results of transcriptome analysis, targeting immune-related genes and reproduction-related genes in the Chinese mitten crab and conducted experiments to further clone the reproduction-related gene by reverse transcript-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods from Chinese mitten crab, compare its sequence with other related proteins. We evaluated the expression of the reproduction-related gene in various tissues by Real-time quantitative reverse transcription-PCR (qRT-PCR), to explore the regulation of reproductive function of these genes during the development. The study is to provide a basis for Chinese mitten crab reproductive mechanisms information, but also provide an important reference for further research on the mechanism of the male reproductive in Chinese mitten crabs. The main results are as follows:
1 Due to its popularity as a traditional food, intensive harvesting of the mitten crab (Eriocheir sinensis) is common, and has lead to an increase in disease incidence, resulting in catastrophic losses to crab aquaculture. The hepatopancreas of E. sinensis is not only an important digestive organ but also an indispensable immune organ. We constructed a non-normalized cDNA library from the hepatopancreas of E. sinensis and acquired 3,297 high-quality expressed sequence tags representing 1,178 unigenes. More than half of these unigenes were novel genes for this species; the remaining had homologs in public databases, which is of great importance for future functional research. We also investigated the association of these genes with immune processes for insight into one of the main functions of the hepatopancreas besides metabolism. Despite the relatively low sampling scalar of our cDNA library, we were able to demonstrate several important properties of the hepatopancreatic transcriptome and identified numerous genes that were closely associated with immune responses. These results might serve as the basis for an in-depth genomics study of E. sinensis, including transcriptome analysis, physical mapping, and whole genome sequencing.
2 Beads with Oligo(dT) are used to isolate poly(A) mRNA after total RNA is collected from accessory sex gland of the Chinese mitten crab. Fragmentation buffer is added for interrupting mRNA to short fragments. Taking these short fragments as templates, random hexamer-primer is used to synthesize the first-strand cDNA. The second-strand cDNA is synthesized using buffer, dNTPs, RNaseH and DNA polymerase I, respectively. Short fragments are purified with QiaQuick PCR extraction kit and resolved with EB buffer for end reparation and adding poly(A). After that, the short fragments are connected with sequencing adapters. And, after the agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates. At last, the library could be sequenced using Illumina HiSeqTM 2000. Transcriptome de novo assembly is carried out with short reads assembling program-SOAPdenovo. A total of 33,221,284 read including total nucleotides 2,657,702,720 nt, which generated 91.06% successful reads. Of these, the average G+C content was 55.19%. These reads were assembled by software consisting of 85,913 contigs, of which 40,977 (47.70%) contigs with the mass of lengths ranging from 100-200nt length. The total length of all the contig is 18,750,722. Functional annotation by Unigene, of which 1713 genes annotated to Article General function prediction only, comment on the Translation, ribosomal structure and biogenesis were 952 genes, associated with the Transcription has 869 Unigene. About Posttranslational modification, protein turnover, chaperones, and Replication, recombination and repair genes were 802 and 789. KEGG is deciphering the genome's database. The accessory sex gland of Chinese mitten crab with 1704 genes (14.99%) in Metabolic pathways, 675 genes involved in Regulation of actin cytoskeleton. Spliceosome has 564 (4.96%) genes involved. Get the original small RNA of fq data, to its connection to low-quality reads, to deal with pollution, access to clean sequence 7939380. Statistics of small RNA (sRNA) species and quantity, and the length distribution of small RNA to do statistics, in general, the length of the small RNA interval 18-30nt, through the length distribution of the peak we can determine the type of small RNA, miRNA concentrated in 21 or 22nt, siRNA concentration in the 24nt, piRNA concentrated in the 30nt. Total Total sRNAs are 7939380, of which miRNA are 382,650 (4.82%), rRNA with 214,013 (2.70%), snRNA with 2573 (0.03%), snoRNA with 563 (0.01%), tRNA with 432,171 (5.44%), unann with 6,907,410 (87.00%). Total Unique sRNAs are 2268484, of which miRNA are 23,021 (1.01%), rRNA with 33,764 (1.49%), snRNA with 1003 (0.04%), snoRNA with 224 (0.01%), tRNA with 29,425 (1.30%), unann with 2,181,047 (96.15%).
3 Leptin is an adipocyte-derived hormone with multiple functions that regulates energy homeostasis and reproductive functions. Increased knowledge of leptin receptor function will enhance our understanding of the physiological roles of leptin in animals. In the present study, a full-length leptin receptor (lepr) cDNA, consisting of 1,353 nucleotides, was cloned from Chinese mitten crab(Eriocheir sinensis) using rapid amplification of cDNA ends (RACE) following the identification of a single expressed sequence tag (EST) clone in a cDNA library. The lepr cDNA consisted of a 22-nucleotide 5'-untranslated region (5'UTR), a 402-nucleotide open reading frame (ORF) and a 929-nucleotide 3'UTR. Multiple sequence alignments revealed that Chinese mitten crab lepr shared a conserved vacuolar protein sorting 55 (Vps55) domain with other species. Chinese mitten crab lepr expression was determined in various tissues and at three different reproductive stages using quantitative real-time RT-PCR. Lepr expression was highest in the intestine, thoracic ganglia, gonad, and accessory gonad, moderate in hepatopancreas and cranial ganglia, and low in muscle, gill, heart, haemocytes, and stomach. Furthermore, lepr expression was significantly higher in the intestine, gonad and thoracic ganglia in immature crabs relative to precocious and mature crabs. In contrast, lepr expression was significantly lower in the hepatopancreas of immature crabs relative to mature crabs. We are the first to identify the lepr gene and to determine its gene expression patterns in various tissues and at three different reproductive stages in Chinese mitten crab. Taken together, our results suggest that lepr may be involved in the nutritional regulation of metabolism and reproduction in Chinese mitten crabs.
4 Ecdysteroid causes various developmental events associated with larval-larval ecdysis. Ecdysteroid-regulated gene could be a putative element of the metamorphic ecdysone response cascade of crustacean. The ecdysteroid-regulated genes mRNA level correlates well with the haemolymph ecdysteroid titre during the molting process. A full-length cDNA of ecdysteroid-regulated gene from Eriocheir sinensis was cloned by the rapid amplification of cDNA ends (RACE), with primers that designed based on the EST sequence of the cDNA library of hepatopancreas from E. sinensis. The cDNA is 1384 bp in size and consists of a 117-bp 5'-untranslated region (UTR), a 306-bp open reading frame (ORF) and an 841-bp 3'-UTR. The genomic DNA is 3059-bp containing two exons interrupted by one intron. Multiple alignments revealed that a conserved Niemann-Pick type C2 domain was identified in the deduced amino acids sequence of ecdysteroid-regulated gene. The mRNA expression of ecdysteroid-regulated gene in ten tissues and the temporal expression in hepatopancreas of four phases were measured by real-time RT-PCR. Ecdysteroid-regulated gene mRNA transcripts could be detected in all examined tissues, and are higher expressed in hepatopancreas than that in other nine tissues (cranial ganglia, P<0.05; the rest eight tissues, P<0.01). Among the four phases, the gene expression at pre-molt is the highest, down-regulated during inter-molt, up-regulated at post-molt, and continues to increase in hard-shell phase. The mRNA expression pattern of ecdysteroid-regulated gene confirmed that the hepatopancreas is a sensitive indicator for ecdysis phase in various crustaceans, and ecdysteroid-regulated gene would be involved in the process of crustacean molting. Therefore, this study by E. sinensis Ecdysteroid-regulated gene cloning and expression analysis, contribute to clarify the mechanism of molting occurs, and may be helpful for the prevention of related problems.
本研究借助分子生物学和生物信息学手段,首次完成对中华绒螯蟹肝胰腺cDNA文库的构建和表达序列标签(EST)的分析,并利用先进的Solexa测序技术,对中华绒螯蟹副性腺组织进行转录组及microRNA的大规模测序并分析其基因表达情况。不仅丰富了中华绒螯蟹的基因组数据库,同时筛选出许多免疫相关及生殖相关的的基因序列。通过对所测高通量数据分析的注释结果,结合已有物种生殖相关基因的研究现状,分析、筛选与该蟹生殖发育相关的部分重要基因；利用RACE和WORKING等技术,获得这些基因的cDNA全长以及基因的结构；利用realtime PCR研究不同基因在不同组织中以及同一组织不同发育阶段的表达情况,探讨这些基因在整个生殖发育过程中的调控作用。这些研究为了解中华绒螯蟹生殖机制提供了基础资料,同时也为深入探讨该蟹雄性生殖机理提供了重要的参考。主要研究结果如下：
1采用常规cDNA文库构建技术,成功构建了一个中华绒螯蟹肝胰腺的EST文库并且获得了3,279高质量序列,代表着1177个一致性序列。超过一半的一致性序列被鉴定是这一物种的未知的新基因。有很大一部分一致性序列(49%)在新发布的数据库中找到它们的同源物,这些同源物对于进一步的功能研究很重要。总结了与代谢和免疫相关的基因,借以考察肝胰腺的两种重要功能：代谢和免疫。为中华绒螯蟹的深入研究打下基础,包括转录组、物理图谱和整个基因组测序。展示了中华绒螯蟹与免疫密切相关的转录组和基因的重要性质。本文重点找到了与机体免疫相关的基因,分为5大类,包括蛋白酶类、抗菌肽、压力蛋白、抗氧化蛋白和其它与免疫相关的基因进行重点分析。蛋白酶类和他们的抑制剂在无脊椎动物抗感染反应中的重要性越来越明显。它们可能在应对侵染性蛋白的体液免役反应过程中表达,这些侵染性蛋白是由侵染性抗原产生的。文库中发现了1种抗菌肽,抗脂多糖因子。压力蛋白
包括铁蛋白、金属硫蛋白、热激蛋白70、胞溶质蛋白A。硒与谷胱甘肽系统在抗氧化防御体系。以及p-1,3-葡聚糖结合蛋白、C型凝集素、血蓝蛋白、干扰素等,都对中华绒螯蟹的免疫有着重要作用。
2从雄性中华绒螯蟹的副性腺组织中提取并分离mRNA,经超声破碎和随机引物反转成cDNA片段,利用Solexa测序(sequencing by synthesis)技术,对这些组织的RNA转录组进行全面测序；用生物信息学的手段和方法,对测序数据进行转录谱分析,分析各组织的基因表达情况,进一步综合各组织之间基因表达的相关性,探讨不同组织在该蟹雄性生殖过程中的作用及调控模式。共获得33,221,284条reads,包含2,657,702,720nt碱基,GC含量55.19%。通过软件组装拼接得到85,913条contig,将所有contig按序列长度从小到大排序,其中40,977条(47.70%) contig集中在100-200nt的长度.从第一条序列开始累加计算contig总长,所有contig的总长度为18,750,722。通过对Unigene的功能注释,其中有1713条基因条注释到General function prediction only,注释上Translation, ribosomal structure and biogenesis有952条基因,与Transcription相关的有869条Unigene。关于Posttranslational modification, protein turnover, chaperones和Replication, recombination and repair的基因分别有802和789条。KEGG是基因组破译方面的数据库。附性腺有1704条基因(14.99%)参与Metabolic pathways,675条基因参与Regulation of actin cytoskeleton。Spliceosome有564(4.96%)条基因参与其中。得到原始小RNA的fq数据后,对其进行去接头,去低质量reads,去污染等处理,得到干净的序列7939380条。统计小RNA(sRNA)的种类(用unique表示)及数量(用total表示),并对小RNA做长度分布统计,一般来说,小RNA的长度区间为18-30nt,通过长度分布的峰我们可以判断小RNA的种类,miRNA集中在21或22nt, siRNA集中在24nt, piRNA集中在30nt。共有Total sRNAs 7939380条,其中miRNA有382650条(4.82%)、rRNA有214013条(2.70%)、snRNA有2573条(0.03%)、snoRNA有563条(0.01%)、rRNA有432171条(5.44%)以及unann有6907410条(87.00%)。共有Unique sRNAs 2268484条,其中miRNA有23021条(1.01%)、rRNA有33764条(1.49%)、snRNA有1003条(0.04%)、snoRNA有224条(0.01%)、tRNA有29425条(1.30%)和unann有2181047条(96.15%)。
3生殖相关蛋白及其功能的研究是当前甲壳动物生殖生物学的研究重点之一,许多生殖相关蛋白及其功能已经明确。瘦素受体在生殖发育过程有调节生殖发育的作用。本研究根据EST提供的信息,采用RACE技术成功克隆了中华绒螯蟹瘦素受体蛋白(EsLEPR)cDNA全长。生物信息学分析发现,该基因cDNA全长序列的长度大小1403 bp包括一个72 bp的5'非翻译区(5' UTR),1929 bp的3'-非翻译区(3'UTR)的一个AATAAA加尾信号和23 bp的polyA尾,和402 bp的开放阅读框(ORF),编码133个氨基酸的多肽与预测分子量14.42kDa和理论等电点为4.46。其基因组DNA序列为3553bp其中有3个内含子和4个外显子组成,包含有典型的(GT和AG)二核苷酸内含子交界处的结构序列剪接位点。氨基酸序列中有一个保守区域：Vacuolar protein sorting 55 (Vps55)。133个氨基酸的序列中的保守区域含有4个跨膜束片段,分别在第4到第26个氨基酸,第33到第52,第67到第89,和第101到第123。氨基酸序列的多重比对结果显示与Eriocheir sinensis同源性最高的是(64%)丽蝇蛹集金小蜂(Nasonia vitripennis)。它与非洲爪蛙(Xenopus laevis,63%)、红鳍东方触(Takifugu rubripes,58%)、人(Homo sapiens,57%)、牛(Bos taurus,57%)、(Rhodnius prolixu,56%)、褐鼠(Rattus norvegicus,55%)、海鞘(Ciona intestinalis,54%),、猕猴(Macaca mulatta,54%),、蚜虫(Acyrthosiphon pisum,48%)和(Boltenia villosa,40%)都有较高的同源性。采用实时定量RT-PCR的方法分析中华绒螯蟹瘦素受体蛋白在不同组织中以(3-actin作为内参的相对表达情况。组织中的表达分析显示,中华绒螯蟹瘦素受体蛋白的mRNA在各个组织中均有表达,主要是在肠、精巢、胸神经节、脑神经节和肝胰腺等组织中表达。
4蜕皮激素引起的幼体蜕皮发育有关的各种活动。蜕皮激素调节蛋白可能是甲壳动物变态蜕皮级联反应的重要元素。在中华绒螯蟹养殖中性早熟和蜕皮未遂的问题经常发生。这两种病害与甲壳动物的蜕皮是密切相关的,研究蜕皮相关基因对于解决中华绒螯蟹性早熟和蜕皮未遂的问题有重要的意义。通过EST分析得到蜕皮激素调节蛋白的两条ESTs (FG358768,FG359773)；用RACE方法得到中华绒螯蟹蜕皮激素调节蛋白cDNA全长。中华绒螯蟹蜕皮激素调节蛋白基因cDNA全长序列的长度大小1384 bp包括一个117-bp的5'非翻译区(5'UTR)、841-bp的3'-非翻译区(3'UTR)的一个AATAAA加尾信号和31-bp的polyA尾,和426 bp的开放阅读框(ORF),编码141个氨基酸的多肽与预测分子量15,150.3 Da和理论等电点为3.86。中华绒螯蟹蜕皮激素调节蛋白的基因组DNA序列为3060bp其中有1个内含子和2个外显子组成。中华绒螯蟹蜕皮激素受体基因组序列的包含有典型的(GT和AG)二核苷酸内含子交界处的结构序列剪接位点。而且在基因组序列序列与其cDNA序列一致。用ClustalW软件对不同物种的蜕皮激素调节蛋白的氨基酸序列进行比对。中华绒螯蟹的蜕皮激素调节蛋白与Litopenaeus vannamei的蜕皮激素调节蛋白(ABD65303)同源性为34.4%,与Bombyx mori蜕皮激素调节蛋白(NP_001134647)的同源性为25.8%。序列比对结果显示有6个高度保守的半胱氨酸残基(Cys22, Cys36, Cys41, Cys87, Cys92, Cys133)。SMART程序分析表明,推导的氨基酸序列包含一个保守的结构域:the Niemann-Pick type C2(属于ML家族)结构域。是由119个氨基酸组成开始在Tyr19并在Val138结束。在ML保守结构域上有22个公认的胆固醇/脂质结合位点。采用SWISS-MODEL模型预测中华绒螯蟹蜕皮激素调节蛋白的三级结构,根据与其他物种相同蛋白很高的相似性,建立的三级结构的模板为:lnepA。模拟结构的氨基酸残基范围开始于第16个残疾和结束于第141残基和序列结构同源性为28.9%。在整个中华绒螯蟹蜕皮激素调节蛋白推导的氨基酸序列结构主要是β-折叠。采用实时定量RT-PCR的方法分析中华绒螯蟹蜕皮激素调节蛋白在不同组织中以β-actin作为内参的相对表达情况。组织中的表达分析显示,中华绒螯蟹蜕皮激素调节蛋白的mRNA主要是在肝胰腺组织中表达,在脑神经节中表达相对较高,在性腺、胃、肌肉和胸神经节中表达较少,而在心,鳃,血淋巴和肠中不表达。在肝胰腺中的表达最丰富,这与肝胰腺的功能密切相关,肝胰腺是蜕皮现象的主要指示器官。采用实时定量RT-PCR的方法,分析中华绒螯蟹蜕皮激素调节蛋白在中华绒螯蟹不同蜕皮时期(蜕皮前、蜕皮中、蜕皮后和硬壳期)肝胰腺中的表达情况。中华绒螯蟹蜕皮激素调节蛋白在蜕皮前的肝胰腺中表达量最高,在进入蜕皮时显著降低,蜕皮后开始回升,直到硬皮期回升到正常水平。
The mitten crab (Eriocheir sinensis) (Henri Milne Edwards 1854), which belongs to Crustacea, Decapoda, Grapsidae, Eriocheir, is a unique species of crab that is native to China. This species is a traditional savory food and therefore comprises one of the economical aquatic species of China. The farming of E. sinensis has been developing rapidly in China over the last two decades and reached its highest yield of 4.0 x 105 tonnes in 2005. With the development of intensive culture, various diseases caused by bacteria, viruses, or rickettsia-like organisms and the sexual precocity, individual small, germplasm degradation are frequent in cultured species populations and have caused catastrophic losses. It has become a major bottleneck in the development of aquaculture industry. Understanding the immune defence mechanisms of the species; characterizing of immune effectors, and the study of reproductive biology, especially the molecular mechanism of reproductive are believed to be helpful in health management and disease control in crab aquaculture, and provide a theoretical to the reproductive control of the artificial crab breeding process.
In the present study, we presented the methodology and results of transcriptome analysis, targeting immune-related genes and reproduction-related genes in the Chinese mitten crab and conducted experiments to further clone the reproduction-related gene by reverse transcript-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods from Chinese mitten crab, compare its sequence with other related proteins. We evaluated the expression of the reproduction-related gene in various tissues by Real-time quantitative reverse transcription-PCR (qRT-PCR), to explore the regulation of reproductive function of these genes during the development. The study is to provide a basis for Chinese mitten crab reproductive mechanisms information, but also provide an important reference for further research on the mechanism of the male reproductive in Chinese mitten crabs. The main results are as follows:
1 Due to its popularity as a traditional food, intensive harvesting of the mitten crab (Eriocheir sinensis) is common, and has lead to an increase in disease incidence, resulting in catastrophic losses to crab aquaculture. The hepatopancreas of E. sinensis is not only an important digestive organ but also an indispensable immune organ. We constructed a non-normalized cDNA library from the hepatopancreas of E. sinensis and acquired 3,297 high-quality expressed sequence tags representing 1,178 unigenes. More than half of these unigenes were novel genes for this species; the remaining had homologs in public databases, which is of great importance for future functional research. We also investigated the association of these genes with immune processes for insight into one of the main functions of the hepatopancreas besides metabolism. Despite the relatively low sampling scalar of our cDNA library, we were able to demonstrate several important properties of the hepatopancreatic transcriptome and identified numerous genes that were closely associated with immune responses. These results might serve as the basis for an in-depth genomics study of E. sinensis, including transcriptome analysis, physical mapping, and whole genome sequencing.
2 Beads with Oligo(dT) are used to isolate poly(A) mRNA after total RNA is collected from accessory sex gland of the Chinese mitten crab. Fragmentation buffer is added for interrupting mRNA to short fragments. Taking these short fragments as templates, random hexamer-primer is used to synthesize the first-strand cDNA. The second-strand cDNA is synthesized using buffer, dNTPs, RNaseH and DNA polymerase I, respectively. Short fragments are purified with QiaQuick PCR extraction kit and resolved with EB buffer for end reparation and adding poly(A). After that, the short fragments are connected with sequencing adapters. And, after the agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates. At last, the library could be sequenced using Illumina HiSeqTM 2000. Transcriptome de novo assembly is carried out with short reads assembling program-SOAPdenovo. A total of 33,221,284 read including total nucleotides 2,657,702,720 nt, which generated 91.06% successful reads. Of these, the average G+C content was 55.19%. These reads were assembled by software consisting of 85,913 contigs, of which 40,977 (47.70%) contigs with the mass of lengths ranging from 100-200nt length. The total length of all the contig is 18,750,722. Functional annotation by Unigene, of which 1713 genes annotated to Article General function prediction only, comment on the Translation, ribosomal structure and biogenesis were 952 genes, associated with the Transcription has 869 Unigene. About Posttranslational modification, protein turnover, chaperones, and Replication, recombination and repair genes were 802 and 789. KEGG is deciphering the genome's database. The accessory sex gland of Chinese mitten crab with 1704 genes (14.99%) in Metabolic pathways, 675 genes involved in Regulation of actin cytoskeleton. Spliceosome has 564 (4.96%) genes involved. Get the original small RNA of fq data, to its connection to low-quality reads, to deal with pollution, access to clean sequence 7939380. Statistics of small RNA (sRNA) species and quantity, and the length distribution of small RNA to do statistics, in general, the length of the small RNA interval 18-30nt, through the length distribution of the peak we can determine the type of small RNA, miRNA concentrated in 21 or 22nt, siRNA concentration in the 24nt, piRNA concentrated in the 30nt. Total Total sRNAs are 7939380, of which miRNA are 382,650 (4.82%), rRNA with 214,013 (2.70%), snRNA with 2573 (0.03%), snoRNA with 563 (0.01%), tRNA with 432,171 (5.44%), unann with 6,907,410 (87.00%). Total Unique sRNAs are 2268484, of which miRNA are 23,021 (1.01%), rRNA with 33,764 (1.49%), snRNA with 1003 (0.04%), snoRNA with 224 (0.01%), tRNA with 29,425 (1.30%), unann with 2,181,047 (96.15%).
3 Leptin is an adipocyte-derived hormone with multiple functions that regulates energy homeostasis and reproductive functions. Increased knowledge of leptin receptor function will enhance our understanding of the physiological roles of leptin in animals. In the present study, a full-length leptin receptor (lepr) cDNA, consisting of 1,353 nucleotides, was cloned from Chinese mitten crab(Eriocheir sinensis) using rapid amplification of cDNA ends (RACE) following the identification of a single expressed sequence tag (EST) clone in a cDNA library. The lepr cDNA consisted of a 22-nucleotide 5'-untranslated region (5'UTR), a 402-nucleotide open reading frame (ORF) and a 929-nucleotide 3'UTR. Multiple sequence alignments revealed that Chinese mitten crab lepr shared a conserved vacuolar protein sorting 55 (Vps55) domain with other species. Chinese mitten crab lepr expression was determined in various tissues and at three different reproductive stages using quantitative real-time RT-PCR. Lepr expression was highest in the intestine, thoracic ganglia, gonad, and accessory gonad, moderate in hepatopancreas and cranial ganglia, and low in muscle, gill, heart, haemocytes, and stomach. Furthermore, lepr expression was significantly higher in the intestine, gonad and thoracic ganglia in immature crabs relative to precocious and mature crabs. In contrast, lepr expression was significantly lower in the hepatopancreas of immature crabs relative to mature crabs. We are the first to identify the lepr gene and to determine its gene expression patterns in various tissues and at three different reproductive stages in Chinese mitten crab. Taken together, our results suggest that lepr may be involved in the nutritional regulation of metabolism and reproduction in Chinese mitten crabs.
4 Ecdysteroid causes various developmental events associated with larval-larval ecdysis. Ecdysteroid-regulated gene could be a putative element of the metamorphic ecdysone response cascade of crustacean. The ecdysteroid-regulated genes mRNA level correlates well with the haemolymph ecdysteroid titre during the molting process. A full-length cDNA of ecdysteroid-regulated gene from Eriocheir sinensis was cloned by the rapid amplification of cDNA ends (RACE), with primers that designed based on the EST sequence of the cDNA library of hepatopancreas from E. sinensis. The cDNA is 1384 bp in size and consists of a 117-bp 5'-untranslated region (UTR), a 306-bp open reading frame (ORF) and an 841-bp 3'-UTR. The genomic DNA is 3059-bp containing two exons interrupted by one intron. Multiple alignments revealed that a conserved Niemann-Pick type C2 domain was identified in the deduced amino acids sequence of ecdysteroid-regulated gene. The mRNA expression of ecdysteroid-regulated gene in ten tissues and the temporal expression in hepatopancreas of four phases were measured by real-time RT-PCR. Ecdysteroid-regulated gene mRNA transcripts could be detected in all examined tissues, and are higher expressed in hepatopancreas than that in other nine tissues (cranial ganglia, P<0.05; the rest eight tissues, P<0.01). Among the four phases, the gene expression at pre-molt is the highest, down-regulated during inter-molt, up-regulated at post-molt, and continues to increase in hard-shell phase. The mRNA expression pattern of ecdysteroid-regulated gene confirmed that the hepatopancreas is a sensitive indicator for ecdysis phase in various crustaceans, and ecdysteroid-regulated gene would be involved in the process of crustacean molting. Therefore, this study by E. sinensis Ecdysteroid-regulated gene cloning and expression analysis, contribute to clarify the mechanism of molting occurs, and may be helpful for the prevention of related problems.
引文
1. Paraoan, L., I. Grierson B.E. Maden (2000) Analysis of expressed sequence tags of retinal pigment epithelium:cystatin C is an abundant transcript. Int J Biochem Cell Biol 32(4):417-26.
2. Zhang, L., Z. Bao, J. Cheng, et al. (2007) Fosmid library construction and initial analysis of end sequences in Zhikong scallop (Chlamys farreri). Mar Biotechnol (NY) 9(5):606-12.
3. La Claire, J.W.,2nd (2006) Analysis of expressed sequence tags from the harmful alga, Prymnesium parvum (Prymnesiophyceae, Haptophyta). Mar Biotechnol (NY) 8(5):534-46.
4. Ronaghi, M., M. Uhlen P. Nyren (1998) A sequencing method based on real-time pyrophosphate. Science 281(5375):363,365.
5. Margulies, M., M. Egholm, W.E. Altaian, et al. (2005) Genome sequencing in microfabricated high-density picolitre reactors. Nature 437(7057):376-80.
6. Mi, S., T. Cai, Y. Hu, et al. (2008) Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5'terminal nucleotide. Cell 133(1):116-27.
7. Montgomery, T.A., M.D. Howell, J.T. Cuperus, et al. (2008) Specificity of ARGONAUTE7-miR390 interaction and dual functionality in TAS3 trans-acting siRNA formation. Cell 133(1):128-41.
8. Schones, D.E., K. Cui, S. Cuddapah, et al. (2008) Dynamic regulation of nucleosome positioning in the human genome. Cell 132(5):887-98.
9. Mardis, E.R. (2008) Next-generation DNA sequencing methods. Annu Rev Genomics Hum Genet 9:387-402.
10. Chang, Y., L. Liang, H. Ma, et al. (2008) Microsatellite analysis of genetic diversity and population structure of Chinese mitten crab (Eriocheir sinensis). J Genet Genomics 35(3):171-6.
11. Gai, Y., J. Zhao, L. Song, et al. (2008) A prophenoloxidase from the Chinese mitten crab Eriocheir sinensis:gene cloning, expression and activity analysis. Fish Shellfish Immunol 24(2):156-67.
12. Rudnick, D., K. Hieb, K. Grimmer, et al. (2003) Patterns and processes of biological invasion:The Chinese mitten crab in San Francisco Bay Basic and Applied Ecology 4:249-262.
13. Wang, H., H. Wang, X. Liang, et al. (2006) Stocking models of Chinese mitten crab (Eriocheir japonica sinensis) in Yangtze lakes. Aquaculture 255:456-465.
14. Sui, L., M. Wille, Y. Cheng, et al. (2007) The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae. Aquaculture 273:139-150.
15. Herborg, L.-M., M.G. Bentley, A.S. Clare, et al. (2006) Mating behaviour and chemical communication in the invasive Chinese mitten crab Eriocheir sinensis. Journal of Experimental Marine Biology and Ecology 329:1-10.
16. Torres, G., M. Charmantier-Daures, S. Chifflet, et al. (2007) Effects of long-term exposure to different salinities on the location and activity of Na+-K+-ATPase in the gills of juvenile mitten crab, Eriocheir sinensis. Comp Biochem Physiol A Mol Integr Physiol 147(2):460-5.
17. Li, X., Y. Lei, X. Gao, et al. (2007) Calcium carbonate supersaturation and precipitation in Chinese mitten crab (Eriocheir japonica sinensis) larval ponds in China:Mass mortality, crystal form analysis, and safety saturation index Aquaculture 272:361-369.
18. Chen, D., M. Zhang S. Shrestha (2007) Compositional characteristics and nutritional quality of Chinese mitten crab (Eriocheir sinensis). Food Chemistry 103:1343-1349.
19. Wu, X., Y. Cheng, L. Sui, et al. (2007) Effect of dietary supplementation of phospholipids and highly unsaturated fatty acids on reproductive performance and offspring quality of Chinese mitten crab, Eriocheir sinensis (H. Milne-Edwards), female broodstock. Aquaculture 273:602-613.
20. Wen, X., L. Chen, C. Ai, et al. (2001) Variation in lipid composition of Chinese mitten-handed crab, Eriocheir sinensis during ovarian maturation. Comp Biochem Physiol B Biochem Mol Biol 130(1):95-104.
21. Normant, M., M. Chrobak A. Szaniawska (2002) Energy value and chemical composition (CHN) of the Chinese mitten crab Eriocheir sinensis (Decapoda:Grapsidae) from the Baltic Sea Thermochimica Acta 394:233-237.
22. Jiang, H., Y.M. Cai, L.Q. Chen, et al. (2009) Functional annotation and analysis of expressed sequence tags from the hepatopancreas of mitten crab (Eriocheir sinensis). Mar Biotechnol (NY) 11(3):317-26.
23. Jiang, H., Y. Yin, X. Zhang, et al. (2009) Chasing relationships between nutrition and reproduction:A comparative transcriptome analysis of hepatopancreas and testis from Eriocheir sinensis. Comparative Biochemistry and Physiology Part D:Genomics and Proteomics 4(3):227-234.
24. Shoji, M., S. Chuma, K. Yoshida, et al. (2005) RNA interference during spermatogenesis in mice. Dev Biol 282(2):524-34.
25. Costa, D.S., C.M.C. Menezes T.A.R. Paula (2007) Spermatogenesis in white-lipped peccaries (Tayassu pecari). Animal Reproduction Science 98:322-334.
26. Dakshinamurti, K. (2005) Biotin-a regulator of gene expression. J Nutr Biochem 16(7):419-23.
27. Eddy, E.M. (1998) Regulation of gene expression during spermatogenesis. Semin Cell Dev Biol 9(4):451-7.
28. Qiu, G.F., R.K. Ramachandra, C.E. Rexroad,3rd, et al. (2008) Molecular characterization and expression profiles of cyclin B1, B2 and Cdc2 kinase during oogenesis and spermatogenesis in rainbow trout (Oncorhynchus mykiss). Anim Reprod Sci 105(3-4):209-25.
29. Feng, H.L., E.Q. Tan, L.J. Zheng, et al. (1998) Expression of HSP-70, estrogen and progesterone receptors in normal and subfertile human testicular tissue. Theriogenology 49:370-370.
30. Adly, M.A., H.A. Assaf M.R. Hussein (2008) Heat shock protein 27 expression in the human testis showing normal and abnormal spermatogenesis. Cell Biol Int 32(10):1247-55.
31. Oshima, R., K. Yoshinaga, Y. Ihara-Ohori, et al. (2007) The Bax Inhibitor-1 needs a functional electron transport chain for cell death suppression. FEBS Lett 581(24):4627-32.
32. Bolduc, N., G.N. Lamb, S.G. Cessna, et al. (2007) Modulation of Bax Inhibitor-1 and cytosolic Ca2+ by cytokinins in Nicotiana tabacum cells Biochimie 89:961-971.
33. Lingner, J., T.R. Hughes, A. Shevchenko, et al. (1997) Reverse transcriptase motifs in the catalytic subunit of telomerase. Science 276(5312):561-7.
34. Zhu, J. A.R. Krainer (2000) Pre-mRNA splicing in the absence of an SR protein RS domain. Genes Dev 14(24):3166-78.
35. Baek, K., R.S. Brown, G. Birrane, et al. (2007) Crystal structure of human cyclin K, a positive regulator of cyclin-dependent kinase 9. J Mol Biol 366(2):563-73.
36. Yu, W., R. Ramakrishnan, Y Wang, et al. (2008) Cyclin T1-dependent genes in activated CD4 T and macrophage cell lines appear enriched in HIV-1 co-factors. PLoS One 3(9):e3146.
37. Fujita, M., H. Takeshita H. Sawa (2007) Cyclin E and CDK2 repress the terminal differentiation of quiescent cells after asymmetric division in C. elegans. PLoS One 2(5):e407.
38. Mikosch, T.S.P., A.S.M. Sonnenberg L.J.L.D.V. Griensven (2001) Isolation, Characterization, and Expression Patterns of a DMC1 Homolog from the Basidiomycete Pleurotus ostreatus Fungal Genetics and Biology 33:59-6.
39. Hayase, A., M. Takagi, T. Miyazaki, et al. (2004) A protein complex containing Mei5 and Sae3 promotes the assembly of the meiosis-specific RecA homolog Dmcl. Cell 119(7):927-40.
40. DeLuca, J.G., Y. Dong, P. Hergert, et al. (2005) Heel and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites. Mol Biol Cell 16(2):519-31.
41. Ramey, C.J., S. Howar, M. Adkins, et al. (2004) Activation of the DNA damage checkpoint in yeast lacking the histone chaperone anti-silencing function 1. Mol Cell Biol 24(23):10313-27.
42. Hashimoto, H., T. Sudo, Y. Mikami, et al. (2008) Germ cell specific protein VASA is over-expressed in epithelial ovarian cancer and disrupts DNA damage-induced G2 checkpoint Gynecologic Oncology 111: 312-319.
43. Caruthers, J.M. D.B. McKay (2002) Helicase structure and mechanism. Curr Opin Struct Biol 12(1):123-33.
44. Skandhan, K.P. A. Rajahariprasad (2007) The process of spermatogenesis liberates significant heat and the scrotum has a role in body thermoregulation. Medical Hypotheses 68:303-307
45. Wu, Q.Y., F. Li, W.J. Zhu, et al. (2007) Cloning, expression, purification, and characterization of arginine kinase from Locusta migratoria manilensis. Comp Biochem Physiol B Biochem Mol Biol 148(4):355-62.
46. Lahnsteiner, F. (2007) Characterization of seminal plasma proteins stabilizing the sperm viability in rainbow trout (Oncorhynchus mykiss). Animal Reproduction Science 97:151-164.
47. Cho, S.Y., M. Chung, M. Park, et al. (2008) ZIFIBI:Prediction of DNA binding sites for zinc finger proteins. Biochem Biophys Res Commun 369(3):845-8.
48. Cecilia, D.A., C. Piero, P. Luisa, et al. (2006) Receptors for leptin and estrogen in the subcommissural organ of rabbits are differentially modulated by fasting Brain Research 1124:62-69.
49. Paz-Filho, GJ., T. Babikian, R. Asarnow, et al. (2008) Leptin Replacement Improves Cognitive Development. PLoS ONE 3(8):e3098.
50. Morash, B.A., A. Imran, D. Wilkinson, et al. (2003) Leptin receptors are developmentally regulated in rat pituitary and hypothalamus Molecular and Cellular Endocrinology 210:1-8.
51. Belgareh-Touze, N., S. Avaro, Y. Rouille, et al. (2002) Yeast Vps55p, a functional homo log of human obesity receptor gene-related protein, is involved in late endosome to vacuole trafficking. Mol Biol Cell 13(5):1694-708.
52. Chen.DW, M. Zhang S. Shrestha (2007) Compositional characteristics and nutritional quality of Chinese mitten crab (Eriocheir sinensis). Food Chemistry 103(4):1343-9.
53. Li, C., J. Zhao, L. Song, et al. (2008) Molecular cloning, genomic organization and functional analysis of an anti-lipopolysaccharide factor from Chinese mitten crab Eriocheir sinensis. Dev Comp Immunol 32(7):784-94.
54. Li, E., L. Chen, C. Zeng, et al. (2008) Comparison of digestive and antioxidant enzymes activities, haemolymph oxyhemocyanin contents and hepatopancreas histology of white shrimp, Litopenaeus vannamei, at various salinities Aquaculture 274(1):80-6.
55. Borkovic, S.S., S.Z. Pavlovic, T.B. Kovacevic, et al. (2008) Antioxidant defence enzyme activities in hepatopancreas, gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube. Comp Biochem Physiol C Toxicol Pharmacol 147(1):122-8.
56. Pinho, G.L., CM. da Rosa, F.E. Maciel, et al. (2005) Antioxidant responses and oxidative stress after microcystin exposure in the hepatopancreas of an estuarine crab species. Ecotoxicol Environ Saf 61(3):353-60.
57. Roux, M.M., A. Pain, K.R. Klimpel, et al. (2002) The lipopolysaccharide and beta-1,3-glucan binding protein gene is upregulated in white spot virus-infected shrimp (Penaeus stylirostris). J Virol 76(14):7140-9.
58. Ried, C., C. Wahl, T. Miethke, et al. (1996) High affinity endotoxin-binding and neutralizing peptides based on the crystal structure of recombinant Limulus anti-lipopolysaccharide factor. J Biol Chem 271(45):28120-7.
59. Sekine, H., A. Kenjo, K. Azumi, et al. (2001) An ancient lectin-dependent complement system in an ascidian:novel lectin isolated from the plasma of the solitary ascidian, Halocynthia roretzi. J Immunol 167(8):4504-10.
60. von Schalburg, K.R., J. Leong, G.A. Cooper, et al. (2008) Rainbow Smelt (Osmerus mordax) Genomic Library and EST Resources. Mar Biotechnol (NY) 10(5):487-91.
61. Douglas, S.E., L.C. Knickle, J. Kimball, et al. (2007) Comprehensive EST analysis of Atlantic halibut (Hippoglossus hippoglossus), a commercially relevant aquaculture species. BMC Genomics 8:144.
62. Sarropoulou, E., D. Nousdili, A. Magoulas, et al. (2008) Linking the genomes of nonmodel teleosts through comparative genomics. Mar Biotechnol (NY) 10(3):227-33.
63. Fujiwara, S., Y. Hirokawa, Y. Takatsuka, et al. (2007) Gene expression profiling of coccolith-bearing cells and naked cells in haptophyte Pleurochrysis haptonemofera with a cDNA macroarray system. Mar Biotechnol (NY) 9(5):550-60.
64. Kocabas, A.M., P. Li, D. Cao, et al. (2002) Expression profile of the channel catfish spleen:analysis of genes involved in immune functions. Mar Biotechnol (NY) 4(6):526-36.
65. Abernathy, J.W., P. Xu, P. Li, et al. (2007) Generation and analysis of expressed sequence tags from the ciliate protozoan parasite Ichthyophthirius multifiliis. BMC Genomics 8:176.
66. Wynne, J.W., M.G. O'Sullivan, M.T. Cook, et al. (2008) Transcriptome analyses of amoebic gill disease-affected Atlantic salmon (Salmo salar) tissues reveal localized host gene suppression. Mar Biotechnol (NY) 10(4):388-403.
67. Wang, Y. X. Guo (2007) Development and characterization of EST-SSR markers in the eastern oyster Crassostrea virginica. Mar Biotechnol (NY) 9(4):500-11.
68. Reusch, T.B., A.S. Veron, C. Preuss, et al. (2008) Comparative analysis of expressed sequence tag (EST) libraries in the seagrass Zostera marina subjected to temperature stress. Mar Biotechnol (NY) 10(3):297-309.
69. Quilang, J., S. Wang, P. Li, et al. (2007) Generation and analysis of ESTs from the eastern oyster, Crassostrea virginica Gmelin and identification of microsatellite and SNP markers. BMC Genomics 8:157.
70. Haidle, L., J.E. Janssen, K. Gharbi, et al. (2008) Determination of Quantitative Trait Loci (QTL) for Early Maturation in Rainbow Trout (Oncorhynchus mykiss). Mar Biotechnol (NY) 10(5):579-92.
71. Ewing, B. P. Green (1998) Base-calling of automated sequencer traces using phred. Ⅱ. Error probabilities. Genome Res 8(3):186-94.
72. Ewing, B., L. Hillier, M.C. Wendl, et al. (1998) Base-calling of automated sequencer traces using phred. Ⅰ. Accuracy assessment. Genome Res 8(3):175-85.
73. Ashburner, M., C.A. Ball, J.A. Blake, et al. (2000) Gene ontology:tool for the unification of biology. The Gene Ontology Consortium. Nat Genet 25(1):25-9.
74. Vizcaino, J.A., F.J. Gonzalez, M.B. Suarez, et al. (2006) Generation, annotation and analysis of ESTs from Trichoderma harzianum CECT 2413. BMC Genomics 7:193.
75. Muhlia-Almazan, A., A. Sanchez-Paz F.L. Garcia-Carreno (2008) Invertebrate trypsins:a review. J Comp Physiol [B].
76. Kim, S.K., P.J. Park, J.B. Kim, et al. (2002) Purification and characterization of a collagenolytic protease from the filefish, Novoden modestrus. J Biochem Mol Biol 35(2):165-71.
77. Hagner-Holler, S., K. Kusche, A. Hembach, et al. (2005) Biochemical and molecular characterisation of hemocyanin from the amphipod Gammarus roeseli: complex pattern of hemocyanin subunit evolution in Crustacea. J Comp Physiol [B] 175(6):445-52.
78. Verri, T., A. Mandal, L. Zilli, et al. (2001) D-glucose transport in decapod crustacean hepatopancreas. Comp Biochem Physiol A Mol Integr Physiol 130(3):585-606.
79. Hoffmann, J.A., F.C. Kafatos, C.A. Janeway, et al. (1999) Phylogenetic perspectives in innate immunity. Science 284(5418):1313-8.
80. Gueguen, Y., J.P. Cadoret, D. Flament, et al. (2003) Immune gene discovery by expressed sequence tags generated from hemocytes of the bacteria-challenged oyster, Crassostrea gigas. Gene 303:139-45.
81. Dong, B. J.H. Xiang (2007) Discovery of genes involved in defense/immunity functions in a haemocytes cDNA library from Fenneropenaeus chinensis by ESTs annotation. Aquaculture 272:208-215.
82. Kam, C.M., M.G. Gotz, G. Koot, et al. (2004) Design and evaluation of inhibitors for dipeptidyl peptidase I (Cathepsin C). Arch Biochem Biophys 427(2):123-34.
83. Staszczak, M. (2008) The role of the ubiquitin-proteasome system in the response of the ligninolytic fungus Trametes versicolor to nitrogen deprivation. Fungal Genet Biol 45(3):328-37.
84. de la Vega, E., N.A. O'Leary, J.E. Shockey, et al. (2008) Anti-lipopolysaccharide factor in Litopenaeus vannamei (LvALF):a broad spectrum antimicrobial peptide essential for shrimp immunity against bacterial and fungal infection. Mol Immunol 45(7):1916-25.
85. Tharntada, S., K. Somboonwiwat, V. Rimphanitchayakit, et al. (2008) Anti-lipopolysaccharide factors from the black tiger shrimp, Penaeus monodon, are encoded by two genomic loci. Fish Shellfish Immunol 24(1):46-54.
86. Lofgren, S.E., L.C. Miletti, M. Steindel, et al. (2008) Trypanocidal and leishmanicidal activities of different antimicrobial peptides (AMPs) isolated from aquatic animals. Exp Parasitol 118(2):197-202.
87. Imjongjirak, C., P. Amparyup, A. Tassanakajon, et al. (2007) Antilipopolysaccharide factor (ALF) of mud crab Scylla paramamosain:molecular cloning, genomic organization and the antimicrobial activity of its synthetic LPS binding domain. Mol Immunol 44(12):3195-203.
88. Nagoshi, H., H. Inagawa, K. Morii, et al. (2006) Cloning and characterization of a LPS-regulatory gene having an LPS binding domain in kuruma prawn Marsupenaeus japonicus. Mol Immunol 43(13):2061-9.
89. Missirlis, F., S. Kosmidis, T. Brody, et al. (2007) Homeostatic mechanisms for iron storage revealed by genetic manipulations and live imaging of Drosophila ferritin. Genetics 177(1):89-100.
90. Larade, K. K.B. Storey (2004) Accumulation and translation of ferritin heavy chain transcripts following anoxia exposure in a marine invertebrate. J Exp Biol 207(Pt 8):1353-60.
91. Brouwer, M., T. Hoexum-Brouwer R.E. Cashon (1993) A putative glutathione-binding site in CdZn-metallothionein identified by equilibrium binding and molecular-modelling studies. Biochem J 294 (Pt 1):219-25.
92. English, T.E. K.B. Storey (2003) Freezing and anoxia stresses induce expression of metallothionein in the foot muscle and hepatopancreas of the marine gastropod Littorina littorea. J Exp Biol 206(Pt 14):2517-24.
93. Multhoff, G. (2007) Heat shock protein 70 (Hsp70):membrane location, export and immunological relevance. Methods 43(3):229-37.
94. Wisen, S. J.E. Gestwicki (2008) Identification of small molecules that modify the protein folding activity of heat shock protein 70. Anal Biochem 374(2):371-7.
95. Yeh, H.Y. P.H. Klesius (2008) Channel catfish, Ictalurus punctatus, cyclophilin A and B cDNA characterization and expression analysis. Vet Immunol Immunopathol 121(3-4):370-7.
96. Zheng, K., P.G Board, X. Fei, et al. (2008) A novel selenium-containing glutathione transferase zetal-1, the activity of which surpasses the level of some native glutathione peroxidases. Int J Biochem Cell Biol.
97. Padhi, A. B. Verghese (2008) Detecting molecular adaptation at individual codons in the pattern recognition protein, lipopolysaccharide-and beta-1,3-glucan-binding protein of decapods. Fish Shellfish Immunol.
98. Jayaraj, S.S., R. Thiagarajan, M. Arumugam, et al. (2007) Isolation, purification and characterization of β 1,3-glucan binding protein from the plasma of marine mussel Perna viridis,. Fish and Shelfish Immunology
99. Luo, T., X. Zhang, Z. Shao, et al. (2003) PmAV, a novel gene involved in virus resistance of shrimp Penaeus monodon. FEBS Lett 551(1-3):53-7.
100. Luo, T., F. Li, K. Lei, et al. (2007) Genomic organization, promoter characterization and expression profiles of an antiviral gene PmAV from the shrimp Penaeus monodon. Mol Immunol 44(7):1516-23.
101. Sun, YD., L.D. Fu, Y.P. Jia, et al. (2008) A hepatopancreas-specific C-type lectin from the Chinese shrimp Fenneropenaeus chinensis exhibits antimicrobial activity. Mol Immunol 45(2):348-61.
102. Vasta, G.R., M. Quesenberry, H. Ahmed, et al. (1999) C-type lectins and galectins mediate innate and adaptive immune functions:their roles in the complement activation pathway. Dev Comp Immunol 23(4-5):401-20.
103. Argraves, W.S., L.M. Greene, M.A. Cooley, et al. (2003) Fibulins:physiological and disease perspectives. EMBO Rep 4(12):1127-31.
104. Clavero-Salas, A., R.R. Sotelo-Mundo, T. Gollas-Galvan, et al. (2007) Transcriptome analysis of gills from the white shrimp Litopenaeus vannamei infected with White Spot Syndrome Virus. Fish Shellfish Immunol 23(2):459-72.
105. Lee, S.Y., B.L. Lee K. Soderhall (2004) Processing of crayfish hemocyanin subunits into phenoloxidase. Biochem Biophys Res Commun 322(2):490-6.
106. Yang, J., Z.L. Wang, X.Q. Zhao, et al. (2008) Natural selection and adaptive evolution of leptin in the ochotona family driven by the cold environmental stress. PLoS One 3(1):e1472.
107. Yuen, B.S., P.C. Owens, M.E. Symonds, et al. (2004) Effects of leptin on fetal plasma adrenocorticotropic hormone and cortisol concentrations and the timing of parturition in the sheep. Biol Reprod 70(6):1650-7.
108. Paula-Lopes, F.F., M. Boelhauve, F.A. Habermann, et al. (2007) Leptin promotes meiotic progression and developmental capacity of bovine oocytes via cumulus cell-independent and-dependent mechanisms. Biol Reprod 76(3):532-41.
109. Paz-Filho, G.J., T. Babikian, R. Asarnow, et al. (2008) Leptin replacement improves cognitive development. PLoS One 3(8):e3098.
110. Kronfeld-Schor, N., J. Zhao, B.A. Silvia, et al. (2000) Steroid-dependent up-regulation of adipose leptin secretion in vitro during pregnancy in mice. Biol Reprod 63(1):274-80.
111. Henson, M.C. V.D. Castracane (2006) Leptin in pregnancy:an update. Biol Reprod 74(2):218-29.
112. Kratzsch, J., M. Hockel W. Kiess (2000) Leptin and pregnancy outcome. Curr Opin Obstet Gynecol 12(6):501-5.
113. Smith, G.D., L.M. Jackson D.L. Foster (2002) Leptin regulation of reproductive function and fertility. Theriogenology 57(1):73-86.
114. Magarinos, M.P., V. Sanchez-Margalet, M. Kotler, et al. (2007) Leptin promotes cell proliferation and survival of trophoblastic cells. Biol Reprod 76(2):203-10.
115. Morash, B.A., A. Imran, D. Wilkinson, et al. (2003) Leptin receptors are developmentally regulated in rat pituitary and hypothalamus. Mol Cell Endocrinol 210(1-2):1-8.
116. Perez-Perez, A., J. Maymo, J.L. Duenas, et al. (2008) Leptin prevents apoptosis of trophoblastic cells by activation of MAPK pathway. Arch Biochem Biophys 477(2):390-5.
117. Huang, Y., K. Ying, Y. Xie, et al. (2001) Cloning and characterization of a novel human leptin receptor overlapping transcript-like 1 gene (LEPROTLl). Biochim Biophys Acta 1517(2):327-31.
118. Rudnick, D.A., K. Hieb, K.F. Grimmer, et al. (2003) Patterns and processes of biological invasion:The Chinese mitten crab in San Francisco Bay. Basic and Applied Ecology 4(3):249-262.
119. Sui, L., M. Wille, Y. Cheng, et al. (2007) The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae. Aquaculture 273(1):139-150.
120. Wang, H.-Z., H.-J. Wang, X.-M. Liang, et al. (2006) Stocking models of Chinese mitten crab (Eriocheir japonica sinensis) in Yangtze lakes. Aquaculture 255(1-4):456-465.
121. Herborg, L.-M., M.G. Bentley, A.S. Clare, et al. (2006) Mating behaviour and chemical communication in the invasive Chinese mitten crab Eriocheir sinensis. Journal of Experimental Marine Biology and Ecology 329(1):1-10.
122. Li, X., Y. Lei, X. Gao, et al. (2007) Calcium carbonate supersaturation and precipitation in Chinese mitten crab (Eriocheir japonica sinensis) larval ponds in China:Mass mortality, crystal form analysis, and safety saturation index. Aquaculture 272(1-4):361-369.
123. Wen, X.B., L.Q. Chen, Z.L. Zhou, et al. (2002) Reproduction response of Chinese mitten-handed crab (Eriocheir sinensis) fed different sources of dietary lipid. Comparative Biochemistry and Physiology-Part A:Molecular & Integrative Physiology 131(3):675-681.
124. Kalinina, M.V., N.A. Vinnikova E.G. Semen'kova (2008) [Gonadogenesis and color characteristics of ovaries in Japanese mitten crab Eriocheir japonicus]. Ontogenez 39(1):58-65.
125. Wheelan, S.J., D.M. Church J.M. Ostell (2001) Spidey:a tool for mRNA-to-genomic alignments. Genome Res 11(11):1952-7.
126. Churbanov, A., M. Pauley, D. Quest, et al. (2005) A method of precise mRNA/DNA homology-based gene structure prediction. BMC Bioinformatics 6:261.
127. Assumpc-a-o, T.C.F., I.M.B. Francischetti, J.F. Andersen, et al. (2008) An insight into the sialome of the blood-sucking bug Triatoma infestans, a vector of Chagas' disease. Insect Biochemistry and Molecular Biology 38(2):213-232.
128. Kumar, S., K. Tamura M. Nei (2004) MEGA3:Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5(2):150-63.
129. Wang, Y, K.K. Kuropatwinski, D.W. White, et al. (1997) Leptin receptor action in hepatic cells. J Biol Chem 272(26):16216-23.
130. Cheng, S.P., C.W. Chi, C.Y. Tzen, et al. (2009) Clinicopathologic significance of leptin and leptin receptor expressions in papillary thyroid carcinoma. Surgery.
131. Zhao, J., L. Song, C. Li, et al. (2007) Molecular cloning, expression of a big defensin gene from bay scallop Argopecten irradians and the antimicrobial activity of its recombinant protein. Mol Immunol 44(4):360-8.
132. Fang, J.J. G.F. Qiu (2009) Molecular cloning of cyclin B transcript with an unusually long 3' untranslation region and its expression analysis during oogenesis in the Chinese mitten crab, Eriocheir sinensis. Mol Biol Rep 36(6):1521-9.
133. Mollineau, W.M., A.O. Adogwa G.W. Garcia (2009) The gross and micro anatomy of the accessory sex glands of the male agouti (Dasyprocta leporina). Anat Histol Embryol 38(3):204-7.
134. Trinadha Babu, B., K. Shyamasundari K. Hanumantha Rao (1989) Studies of the neurohaemal organ (sinus gland) of the portunid crab Portunus sanguinolentus (Herbst) (Brachyura).1. Morphology and histology in relation to moulting. Folia Morphol (Praha) 37(4):388-93.
135. Zarubin, T.P., E.S. Chang D.L. Mykles (2009) Expression of recombinant eyestalk crustacean hyperglycemic hormone from the tropical land crab, Gecarcinus lateralis, that inhibits Y-organ ecdysteroidogenesis in vitro. Mol Biol Rep 36(6):1231-7.
136. Normant, M., M. Chrobak A. Szaniawska (2002) Energy value and chemical composition (CHN) of the Chinese mitten crab Eriocheir sinensis (Decapoda:Grapsidae) from the Baltic Sea. Thermochimica Acta 394(1-2):233-237.
137. Yamaguchi, S., F.T. Celino, A. Ito, et al. (2008) Effects of arsenic on gonadal development in freshwater crab, Somanniathelphusa pax, in Vietnam and Geothelphusa dehaani in Japan. Ecotoxicology 17(8):772-80.
138. Henault, M.A., G.J. Killian, J.F. Kavanaugh, et al. (1995) Effect of accessory sex gland fluid from bulls of differing fertilities on the ability of cauda epididymal sperm to penetrate zona-free bovine oocytes. Biol Reprod 52(2):390-7.
139. Abd el-Hamid, N.F. (1988) Size composition, sex ratio, size at maturity, mating and egg-laying in the crab Portunus pelagicus (Linnaeus). Folia Morphol (Praha) 36(2):200-10.
140. Le Bizec, B., J.-P. Antignac, F. Monteau, et al. (2002) Ecdysteroids:one potential new anabolic family in breeding animals. Analytica Chimica Acta 473(1-2):89-97.
141. Margam, V.M., D.B. Gelman S.R. Palli (2006) Ecdysteroid titers and developmental expression of ecdysteroid-regulated genes during metamorphosis of the yellow fever mosquito, Aedes aegypti (Diptera:Culicidae). Journal of Insect Physiology 52(6):558-568.
142. Burtis, K.C., C.S. Thummel, C.W. Jones, et al. (1990) The Drosophila 74EF early puff contains E74, a complex ecdysone-inducible gene that encodes two ets-related proteins. Cell 61(1):85-99.
143. Sekimoto, T., M. Iwami S. Sakurai (2007) 20-Hydroxyecdysone regulation of two isoforms of the Ets transcription factor E74 gene in programmed cell death in the silkworm anterior silk gland. Insect Mol Biol 16(5):581-90.
144. Hepperle, C. K. Hartfelder (2001) Differentially expressed regulatory genes in honey bee caste development. Naturwissenschaften 88(3):113-6.
145. Pongs, O. (1988) Ecdysteroid-regulated gene expression in Drosophila melanogaster. Eur J Biochem 175(2):199-204.
146. Paul, R.K., H. Takeuchi, Y. Matsuo, et al. (2005) Gene expression of ecdysteroid-regulated gene E74 of the honeybee in ovary and brain. Insect Mol Biol 14(1):9-15.
147. Mu, X.Y. G.A. Leblanc (2004) Cross communication between signaling pathways:Juvenoid hormones modulate ecdysteroid activity in a crustacean. Journal of Experimental Zoology Part a-Comparative Experimental Biology 301A(10):793-801.
148. Matsuoka, T. H. Fujiwara (2000) Expression of ecdysteroid-regulated genes is reduced specifically in the wing discs of the wing-deficient mutant (fl) of Bombyx mori. Development Genes and Evolution 210(3):120-128.
149. Arnold, K., L. Bordoli, J. Kopp, et al. (2006) The SWISS-MODEL workspace:a web-based environment for protein structure homology modelling. Bioinformatics 22(2):195-201.
150. Schwede, T., J. Kopp, N. Guex, et al. (2003) SWISS-MODEL:An automated protein homology-modeling server. Nucleic Acids Res 31(13):3381-5.
151. Guex, N. M.C. Peitsch (1997) SWISS-MODEL and the Swiss-PdbViewer:an environment for comparative protein modeling. Electrophoresis 18(15):2714-23.
152. Liou, H.L., S.S. Dixit, S. Xu, et al. (2006) NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. J Biol Chem 281 (48):36710-23.
153. Maroy, P., R. Dennis, C. Beckers, et al. (1978) Demonstration of an ecdysteroid receptor in a cultured cell line of Drosophila melanogaster. Proc Natl Acad Sci U S A 75(12):6035-8.
154. Yund, M.A., D.S. King J.W. Fristrom (1978) Ecdysteroid receptors in imaginal discs of Drosophila melanogaster. Proc Natl Acad Sci U S A 75(12):6039-43.
155. Lan, Q., Z. Wu L.M. Riddiford (1997) Regulation of the ecdysone receptor, USP, E75 and MHR3 mRNAs by 20-hydroxyecdysone in the GVI cell line of the tobacco hornworm, Manduca sexta. Insect Mol Biol 6(1):3-10.
156. Barker, G.C. H.H. Rees (1990) Ecdysteroids in nematodes. Parasitol Today 6(12):384-7.
157. Mathias, J.D., Y. Ran, J.D. Carter, et al. (2009) Interactions of the GM2 activator protein with phosphatidylcholine bilayers:a site-directed spin-labeling power saturation study. Biophys J 97(5):1436-44.
158. Jelinek, D., S.M. Patrick, K.N. Kitt, et al. (2009) Physiological and coordinate downregulation of the NPC1 and NPC2 genes are associated with the sequestration of LDL-derived cholesterol within endocytic compartments. J Cell Biochem 108(5):1102-16.
159. Record, E., S. Moukha M. Asther (1999) Characterization and expression of the cDNA encoding a new kind of phospholipid transfer protein, the phosphatidylglycerol/phosphatidylinositol transfer protein from Aspergillus oryzae:evidence of a putative membrane targeted phospholipid transfer protein in fungi. Biochim Biophys Acta 1444(2):276-82.
160. Ichikawa, S., T. Takai, T. Yashiki, et al. (2009) Lipopolysaccharide binding of the mite allergen Der f 2. Genes Cells 14(9):1055-65.
161. Park, S.Y., J.H. Cho, D.Y. Oh, et al. (2009) House dust mite allergen Der f 2-induced phospholipase Dl activation is critical for the production of interleukin-13 through activating transcription factor-2 activation in human bronchial epithelial cells. J Biol Chem 284(30):20099-110.
162. Xie, B., W. Wang D.J. Mahuran (1992) A Cys138-to-Arg substitution in the GM2 activator protein is associated with the AB variant form of GM2 gangliosidosis. Am J Hum Genet 50(5):1046-52.
163. Elleder, M., H. Houstkova, J. Zeman, et al. (2001) Pulmonary storage with emphysema as a sign of Niemann-Pick type C2 disease (second complementation group). Report of a case. Virchows Arch 439(2):206-11.
164. Xu, S., B. Benoff, H.L. Liou, et al. (2007) Structural basis of sterol binding by NPC2, a lysosomal protein deficient in Niemann-Pick type C2 disease. J Biol Chem 282(32):23525-31.
165. Babalola, J.O., M. Wendeler, B. Breiden, et al. (2007) Development of an assay for the intermembrane transfer of cholesterol by Niemann-Pick C2 protein. Biol Chem 388(6):617-26.
166. Busso, D., MJ. Onate-Alvarado, E. Balboa, et al. (2009) Female infertility due to anovulation and defective steroidogenesis in NPC2 deficient mice. Mol Cell Endocrinol.
167. Friedland, N., H.L. Liou, P. Lobel, et al. (2003) Structure of a cholesterol-binding protein deficient in Niemann-Pick type C2 disease. Proc Natl Acad Sci U S A 100(5):2512-7.
168. Kataoka, H., H. Nagasawa, A. Isogai, et al. (1991) Prothoracicotropic hormone of the silkworm, Bombyx mori:amino acid sequence and dimeric structure. Agric Biol Chem 55(1):73-86.
169. Marchal, E., H.P. Vandersmissen, L. Badisco, et al. (2009) Control of ecdysteroidogenesis in prothoracic glands of insects:a review. Peptides 31(3):506-19.
170. Covi, J.A., B.D. Bader, E.S. Chang, et al. Molt cycle regulation of protein synthesis in skeletal muscle of the blackback land crab, Gecarcinus lateralis, and the differential expression of a myostatin-like factor during atrophy induced by molting or unweighting. J Exp Biol 213(Pt1):172-83.
171. Mykles, D.L. D.M. Skinner (1982) Molt cycle-associated changes in calcium-dependent proteinase activity that degrades actin and myosin in crustacean muscle. Dev Biol 92(2):386-97.
172. Zmora, N., J. Trant, Y. Zohar, et al. (2009) Mo It-inhibiting hormone stimulates vitellogenesis at advanced ovarian developmental stages in the female blue crab, Callinectes sapidus 1:an ovarian stage dependent involvement. Saline Systems 5:7.
173. Reddy, P.S., A. Bhagylakshmi R. Ramamurthi (1985) Molt-inhibition in the crab Oziotelphusa senex senex following exposure to malathion and methyl parathion. Bull Environ Contam Toxicol 35(1):92-7.
174. Han, D.W., N. Patel R. Douglas Watson (2006) Regulation of protein synthesis in Y-organs of the blue crab (Callinectes sapidus):involvement of cyclic AMP. J Exp Zool A Comp Exp Biol 305(4):328-34.
175. Spindler, K.D., D. Adelung C. Tchernigovtzeff (1974) A comparison of the methods of molt staging according to Drach and to Adelung in the common shore crab, Carcinus maenas. Z Naturforsch C 29(11-12):754-6.
176. Nakatsuji, T., C.Y. Lee R.D. Watson (2009) Crustacean molt-inhibiting hormone:structure, function, and cellular mode of action. Comp Biochem Physiol A Mol Integr Physiol 152(2):139-48.
177. Subramoniam, T. (2000) Crustacean ecdysteriods in reproduction and embryogenesis. Comp Biochem Physiol C Toxicol Pharmacol 125(2):135-56.
178. Meszaros, M. D.B. Morton (1997) Up-and downregulation of esr20, an ecdysteroid-regulated gene expressed in the tracheae of Manduca sexta. Arch Insect Biochem Physiol 34(2):159-74.
179. Paul, R.K., H. Takeuchi T. Kubo (2006) Expression of two ecdysteroid-regulated genes, Broad-Complex and E75, in the brain and ovary of the honeybee (Apis mellifera L.). Zoolog Sci 23(12):1085-92.
2. Zhang, L., Z. Bao, J. Cheng, et al. (2007) Fosmid library construction and initial analysis of end sequences in Zhikong scallop (Chlamys farreri). Mar Biotechnol (NY) 9(5):606-12.
3. La Claire, J.W.,2nd (2006) Analysis of expressed sequence tags from the harmful alga, Prymnesium parvum (Prymnesiophyceae, Haptophyta). Mar Biotechnol (NY) 8(5):534-46.
4. Ronaghi, M., M. Uhlen P. Nyren (1998) A sequencing method based on real-time pyrophosphate. Science 281(5375):363,365.
5. Margulies, M., M. Egholm, W.E. Altaian, et al. (2005) Genome sequencing in microfabricated high-density picolitre reactors. Nature 437(7057):376-80.
6. Mi, S., T. Cai, Y. Hu, et al. (2008) Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5'terminal nucleotide. Cell 133(1):116-27.
7. Montgomery, T.A., M.D. Howell, J.T. Cuperus, et al. (2008) Specificity of ARGONAUTE7-miR390 interaction and dual functionality in TAS3 trans-acting siRNA formation. Cell 133(1):128-41.
8. Schones, D.E., K. Cui, S. Cuddapah, et al. (2008) Dynamic regulation of nucleosome positioning in the human genome. Cell 132(5):887-98.
9. Mardis, E.R. (2008) Next-generation DNA sequencing methods. Annu Rev Genomics Hum Genet 9:387-402.
10. Chang, Y., L. Liang, H. Ma, et al. (2008) Microsatellite analysis of genetic diversity and population structure of Chinese mitten crab (Eriocheir sinensis). J Genet Genomics 35(3):171-6.
11. Gai, Y., J. Zhao, L. Song, et al. (2008) A prophenoloxidase from the Chinese mitten crab Eriocheir sinensis:gene cloning, expression and activity analysis. Fish Shellfish Immunol 24(2):156-67.
12. Rudnick, D., K. Hieb, K. Grimmer, et al. (2003) Patterns and processes of biological invasion:The Chinese mitten crab in San Francisco Bay Basic and Applied Ecology 4:249-262.
13. Wang, H., H. Wang, X. Liang, et al. (2006) Stocking models of Chinese mitten crab (Eriocheir japonica sinensis) in Yangtze lakes. Aquaculture 255:456-465.
14. Sui, L., M. Wille, Y. Cheng, et al. (2007) The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae. Aquaculture 273:139-150.
15. Herborg, L.-M., M.G. Bentley, A.S. Clare, et al. (2006) Mating behaviour and chemical communication in the invasive Chinese mitten crab Eriocheir sinensis. Journal of Experimental Marine Biology and Ecology 329:1-10.
16. Torres, G., M. Charmantier-Daures, S. Chifflet, et al. (2007) Effects of long-term exposure to different salinities on the location and activity of Na+-K+-ATPase in the gills of juvenile mitten crab, Eriocheir sinensis. Comp Biochem Physiol A Mol Integr Physiol 147(2):460-5.
17. Li, X., Y. Lei, X. Gao, et al. (2007) Calcium carbonate supersaturation and precipitation in Chinese mitten crab (Eriocheir japonica sinensis) larval ponds in China:Mass mortality, crystal form analysis, and safety saturation index Aquaculture 272:361-369.
18. Chen, D., M. Zhang S. Shrestha (2007) Compositional characteristics and nutritional quality of Chinese mitten crab (Eriocheir sinensis). Food Chemistry 103:1343-1349.
19. Wu, X., Y. Cheng, L. Sui, et al. (2007) Effect of dietary supplementation of phospholipids and highly unsaturated fatty acids on reproductive performance and offspring quality of Chinese mitten crab, Eriocheir sinensis (H. Milne-Edwards), female broodstock. Aquaculture 273:602-613.
20. Wen, X., L. Chen, C. Ai, et al. (2001) Variation in lipid composition of Chinese mitten-handed crab, Eriocheir sinensis during ovarian maturation. Comp Biochem Physiol B Biochem Mol Biol 130(1):95-104.
21. Normant, M., M. Chrobak A. Szaniawska (2002) Energy value and chemical composition (CHN) of the Chinese mitten crab Eriocheir sinensis (Decapoda:Grapsidae) from the Baltic Sea Thermochimica Acta 394:233-237.
22. Jiang, H., Y.M. Cai, L.Q. Chen, et al. (2009) Functional annotation and analysis of expressed sequence tags from the hepatopancreas of mitten crab (Eriocheir sinensis). Mar Biotechnol (NY) 11(3):317-26.
23. Jiang, H., Y. Yin, X. Zhang, et al. (2009) Chasing relationships between nutrition and reproduction:A comparative transcriptome analysis of hepatopancreas and testis from Eriocheir sinensis. Comparative Biochemistry and Physiology Part D:Genomics and Proteomics 4(3):227-234.
24. Shoji, M., S. Chuma, K. Yoshida, et al. (2005) RNA interference during spermatogenesis in mice. Dev Biol 282(2):524-34.
25. Costa, D.S., C.M.C. Menezes T.A.R. Paula (2007) Spermatogenesis in white-lipped peccaries (Tayassu pecari). Animal Reproduction Science 98:322-334.
26. Dakshinamurti, K. (2005) Biotin-a regulator of gene expression. J Nutr Biochem 16(7):419-23.
27. Eddy, E.M. (1998) Regulation of gene expression during spermatogenesis. Semin Cell Dev Biol 9(4):451-7.
28. Qiu, G.F., R.K. Ramachandra, C.E. Rexroad,3rd, et al. (2008) Molecular characterization and expression profiles of cyclin B1, B2 and Cdc2 kinase during oogenesis and spermatogenesis in rainbow trout (Oncorhynchus mykiss). Anim Reprod Sci 105(3-4):209-25.
29. Feng, H.L., E.Q. Tan, L.J. Zheng, et al. (1998) Expression of HSP-70, estrogen and progesterone receptors in normal and subfertile human testicular tissue. Theriogenology 49:370-370.
30. Adly, M.A., H.A. Assaf M.R. Hussein (2008) Heat shock protein 27 expression in the human testis showing normal and abnormal spermatogenesis. Cell Biol Int 32(10):1247-55.
31. Oshima, R., K. Yoshinaga, Y. Ihara-Ohori, et al. (2007) The Bax Inhibitor-1 needs a functional electron transport chain for cell death suppression. FEBS Lett 581(24):4627-32.
32. Bolduc, N., G.N. Lamb, S.G. Cessna, et al. (2007) Modulation of Bax Inhibitor-1 and cytosolic Ca2+ by cytokinins in Nicotiana tabacum cells Biochimie 89:961-971.
33. Lingner, J., T.R. Hughes, A. Shevchenko, et al. (1997) Reverse transcriptase motifs in the catalytic subunit of telomerase. Science 276(5312):561-7.
34. Zhu, J. A.R. Krainer (2000) Pre-mRNA splicing in the absence of an SR protein RS domain. Genes Dev 14(24):3166-78.
35. Baek, K., R.S. Brown, G. Birrane, et al. (2007) Crystal structure of human cyclin K, a positive regulator of cyclin-dependent kinase 9. J Mol Biol 366(2):563-73.
36. Yu, W., R. Ramakrishnan, Y Wang, et al. (2008) Cyclin T1-dependent genes in activated CD4 T and macrophage cell lines appear enriched in HIV-1 co-factors. PLoS One 3(9):e3146.
37. Fujita, M., H. Takeshita H. Sawa (2007) Cyclin E and CDK2 repress the terminal differentiation of quiescent cells after asymmetric division in C. elegans. PLoS One 2(5):e407.
38. Mikosch, T.S.P., A.S.M. Sonnenberg L.J.L.D.V. Griensven (2001) Isolation, Characterization, and Expression Patterns of a DMC1 Homolog from the Basidiomycete Pleurotus ostreatus Fungal Genetics and Biology 33:59-6.
39. Hayase, A., M. Takagi, T. Miyazaki, et al. (2004) A protein complex containing Mei5 and Sae3 promotes the assembly of the meiosis-specific RecA homolog Dmcl. Cell 119(7):927-40.
40. DeLuca, J.G., Y. Dong, P. Hergert, et al. (2005) Heel and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites. Mol Biol Cell 16(2):519-31.
41. Ramey, C.J., S. Howar, M. Adkins, et al. (2004) Activation of the DNA damage checkpoint in yeast lacking the histone chaperone anti-silencing function 1. Mol Cell Biol 24(23):10313-27.
42. Hashimoto, H., T. Sudo, Y. Mikami, et al. (2008) Germ cell specific protein VASA is over-expressed in epithelial ovarian cancer and disrupts DNA damage-induced G2 checkpoint Gynecologic Oncology 111: 312-319.
43. Caruthers, J.M. D.B. McKay (2002) Helicase structure and mechanism. Curr Opin Struct Biol 12(1):123-33.
44. Skandhan, K.P. A. Rajahariprasad (2007) The process of spermatogenesis liberates significant heat and the scrotum has a role in body thermoregulation. Medical Hypotheses 68:303-307
45. Wu, Q.Y., F. Li, W.J. Zhu, et al. (2007) Cloning, expression, purification, and characterization of arginine kinase from Locusta migratoria manilensis. Comp Biochem Physiol B Biochem Mol Biol 148(4):355-62.
46. Lahnsteiner, F. (2007) Characterization of seminal plasma proteins stabilizing the sperm viability in rainbow trout (Oncorhynchus mykiss). Animal Reproduction Science 97:151-164.
47. Cho, S.Y., M. Chung, M. Park, et al. (2008) ZIFIBI:Prediction of DNA binding sites for zinc finger proteins. Biochem Biophys Res Commun 369(3):845-8.
48. Cecilia, D.A., C. Piero, P. Luisa, et al. (2006) Receptors for leptin and estrogen in the subcommissural organ of rabbits are differentially modulated by fasting Brain Research 1124:62-69.
49. Paz-Filho, GJ., T. Babikian, R. Asarnow, et al. (2008) Leptin Replacement Improves Cognitive Development. PLoS ONE 3(8):e3098.
50. Morash, B.A., A. Imran, D. Wilkinson, et al. (2003) Leptin receptors are developmentally regulated in rat pituitary and hypothalamus Molecular and Cellular Endocrinology 210:1-8.
51. Belgareh-Touze, N., S. Avaro, Y. Rouille, et al. (2002) Yeast Vps55p, a functional homo log of human obesity receptor gene-related protein, is involved in late endosome to vacuole trafficking. Mol Biol Cell 13(5):1694-708.
52. Chen.DW, M. Zhang S. Shrestha (2007) Compositional characteristics and nutritional quality of Chinese mitten crab (Eriocheir sinensis). Food Chemistry 103(4):1343-9.
53. Li, C., J. Zhao, L. Song, et al. (2008) Molecular cloning, genomic organization and functional analysis of an anti-lipopolysaccharide factor from Chinese mitten crab Eriocheir sinensis. Dev Comp Immunol 32(7):784-94.
54. Li, E., L. Chen, C. Zeng, et al. (2008) Comparison of digestive and antioxidant enzymes activities, haemolymph oxyhemocyanin contents and hepatopancreas histology of white shrimp, Litopenaeus vannamei, at various salinities Aquaculture 274(1):80-6.
55. Borkovic, S.S., S.Z. Pavlovic, T.B. Kovacevic, et al. (2008) Antioxidant defence enzyme activities in hepatopancreas, gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube. Comp Biochem Physiol C Toxicol Pharmacol 147(1):122-8.
56. Pinho, G.L., CM. da Rosa, F.E. Maciel, et al. (2005) Antioxidant responses and oxidative stress after microcystin exposure in the hepatopancreas of an estuarine crab species. Ecotoxicol Environ Saf 61(3):353-60.
57. Roux, M.M., A. Pain, K.R. Klimpel, et al. (2002) The lipopolysaccharide and beta-1,3-glucan binding protein gene is upregulated in white spot virus-infected shrimp (Penaeus stylirostris). J Virol 76(14):7140-9.
58. Ried, C., C. Wahl, T. Miethke, et al. (1996) High affinity endotoxin-binding and neutralizing peptides based on the crystal structure of recombinant Limulus anti-lipopolysaccharide factor. J Biol Chem 271(45):28120-7.
59. Sekine, H., A. Kenjo, K. Azumi, et al. (2001) An ancient lectin-dependent complement system in an ascidian:novel lectin isolated from the plasma of the solitary ascidian, Halocynthia roretzi. J Immunol 167(8):4504-10.
60. von Schalburg, K.R., J. Leong, G.A. Cooper, et al. (2008) Rainbow Smelt (Osmerus mordax) Genomic Library and EST Resources. Mar Biotechnol (NY) 10(5):487-91.
61. Douglas, S.E., L.C. Knickle, J. Kimball, et al. (2007) Comprehensive EST analysis of Atlantic halibut (Hippoglossus hippoglossus), a commercially relevant aquaculture species. BMC Genomics 8:144.
62. Sarropoulou, E., D. Nousdili, A. Magoulas, et al. (2008) Linking the genomes of nonmodel teleosts through comparative genomics. Mar Biotechnol (NY) 10(3):227-33.
63. Fujiwara, S., Y. Hirokawa, Y. Takatsuka, et al. (2007) Gene expression profiling of coccolith-bearing cells and naked cells in haptophyte Pleurochrysis haptonemofera with a cDNA macroarray system. Mar Biotechnol (NY) 9(5):550-60.
64. Kocabas, A.M., P. Li, D. Cao, et al. (2002) Expression profile of the channel catfish spleen:analysis of genes involved in immune functions. Mar Biotechnol (NY) 4(6):526-36.
65. Abernathy, J.W., P. Xu, P. Li, et al. (2007) Generation and analysis of expressed sequence tags from the ciliate protozoan parasite Ichthyophthirius multifiliis. BMC Genomics 8:176.
66. Wynne, J.W., M.G. O'Sullivan, M.T. Cook, et al. (2008) Transcriptome analyses of amoebic gill disease-affected Atlantic salmon (Salmo salar) tissues reveal localized host gene suppression. Mar Biotechnol (NY) 10(4):388-403.
67. Wang, Y. X. Guo (2007) Development and characterization of EST-SSR markers in the eastern oyster Crassostrea virginica. Mar Biotechnol (NY) 9(4):500-11.
68. Reusch, T.B., A.S. Veron, C. Preuss, et al. (2008) Comparative analysis of expressed sequence tag (EST) libraries in the seagrass Zostera marina subjected to temperature stress. Mar Biotechnol (NY) 10(3):297-309.
69. Quilang, J., S. Wang, P. Li, et al. (2007) Generation and analysis of ESTs from the eastern oyster, Crassostrea virginica Gmelin and identification of microsatellite and SNP markers. BMC Genomics 8:157.
70. Haidle, L., J.E. Janssen, K. Gharbi, et al. (2008) Determination of Quantitative Trait Loci (QTL) for Early Maturation in Rainbow Trout (Oncorhynchus mykiss). Mar Biotechnol (NY) 10(5):579-92.
71. Ewing, B. P. Green (1998) Base-calling of automated sequencer traces using phred. Ⅱ. Error probabilities. Genome Res 8(3):186-94.
72. Ewing, B., L. Hillier, M.C. Wendl, et al. (1998) Base-calling of automated sequencer traces using phred. Ⅰ. Accuracy assessment. Genome Res 8(3):175-85.
73. Ashburner, M., C.A. Ball, J.A. Blake, et al. (2000) Gene ontology:tool for the unification of biology. The Gene Ontology Consortium. Nat Genet 25(1):25-9.
74. Vizcaino, J.A., F.J. Gonzalez, M.B. Suarez, et al. (2006) Generation, annotation and analysis of ESTs from Trichoderma harzianum CECT 2413. BMC Genomics 7:193.
75. Muhlia-Almazan, A., A. Sanchez-Paz F.L. Garcia-Carreno (2008) Invertebrate trypsins:a review. J Comp Physiol [B].
76. Kim, S.K., P.J. Park, J.B. Kim, et al. (2002) Purification and characterization of a collagenolytic protease from the filefish, Novoden modestrus. J Biochem Mol Biol 35(2):165-71.
77. Hagner-Holler, S., K. Kusche, A. Hembach, et al. (2005) Biochemical and molecular characterisation of hemocyanin from the amphipod Gammarus roeseli: complex pattern of hemocyanin subunit evolution in Crustacea. J Comp Physiol [B] 175(6):445-52.
78. Verri, T., A. Mandal, L. Zilli, et al. (2001) D-glucose transport in decapod crustacean hepatopancreas. Comp Biochem Physiol A Mol Integr Physiol 130(3):585-606.
79. Hoffmann, J.A., F.C. Kafatos, C.A. Janeway, et al. (1999) Phylogenetic perspectives in innate immunity. Science 284(5418):1313-8.
80. Gueguen, Y., J.P. Cadoret, D. Flament, et al. (2003) Immune gene discovery by expressed sequence tags generated from hemocytes of the bacteria-challenged oyster, Crassostrea gigas. Gene 303:139-45.
81. Dong, B. J.H. Xiang (2007) Discovery of genes involved in defense/immunity functions in a haemocytes cDNA library from Fenneropenaeus chinensis by ESTs annotation. Aquaculture 272:208-215.
82. Kam, C.M., M.G. Gotz, G. Koot, et al. (2004) Design and evaluation of inhibitors for dipeptidyl peptidase I (Cathepsin C). Arch Biochem Biophys 427(2):123-34.
83. Staszczak, M. (2008) The role of the ubiquitin-proteasome system in the response of the ligninolytic fungus Trametes versicolor to nitrogen deprivation. Fungal Genet Biol 45(3):328-37.
84. de la Vega, E., N.A. O'Leary, J.E. Shockey, et al. (2008) Anti-lipopolysaccharide factor in Litopenaeus vannamei (LvALF):a broad spectrum antimicrobial peptide essential for shrimp immunity against bacterial and fungal infection. Mol Immunol 45(7):1916-25.
85. Tharntada, S., K. Somboonwiwat, V. Rimphanitchayakit, et al. (2008) Anti-lipopolysaccharide factors from the black tiger shrimp, Penaeus monodon, are encoded by two genomic loci. Fish Shellfish Immunol 24(1):46-54.
86. Lofgren, S.E., L.C. Miletti, M. Steindel, et al. (2008) Trypanocidal and leishmanicidal activities of different antimicrobial peptides (AMPs) isolated from aquatic animals. Exp Parasitol 118(2):197-202.
87. Imjongjirak, C., P. Amparyup, A. Tassanakajon, et al. (2007) Antilipopolysaccharide factor (ALF) of mud crab Scylla paramamosain:molecular cloning, genomic organization and the antimicrobial activity of its synthetic LPS binding domain. Mol Immunol 44(12):3195-203.
88. Nagoshi, H., H. Inagawa, K. Morii, et al. (2006) Cloning and characterization of a LPS-regulatory gene having an LPS binding domain in kuruma prawn Marsupenaeus japonicus. Mol Immunol 43(13):2061-9.
89. Missirlis, F., S. Kosmidis, T. Brody, et al. (2007) Homeostatic mechanisms for iron storage revealed by genetic manipulations and live imaging of Drosophila ferritin. Genetics 177(1):89-100.
90. Larade, K. K.B. Storey (2004) Accumulation and translation of ferritin heavy chain transcripts following anoxia exposure in a marine invertebrate. J Exp Biol 207(Pt 8):1353-60.
91. Brouwer, M., T. Hoexum-Brouwer R.E. Cashon (1993) A putative glutathione-binding site in CdZn-metallothionein identified by equilibrium binding and molecular-modelling studies. Biochem J 294 (Pt 1):219-25.
92. English, T.E. K.B. Storey (2003) Freezing and anoxia stresses induce expression of metallothionein in the foot muscle and hepatopancreas of the marine gastropod Littorina littorea. J Exp Biol 206(Pt 14):2517-24.
93. Multhoff, G. (2007) Heat shock protein 70 (Hsp70):membrane location, export and immunological relevance. Methods 43(3):229-37.
94. Wisen, S. J.E. Gestwicki (2008) Identification of small molecules that modify the protein folding activity of heat shock protein 70. Anal Biochem 374(2):371-7.
95. Yeh, H.Y. P.H. Klesius (2008) Channel catfish, Ictalurus punctatus, cyclophilin A and B cDNA characterization and expression analysis. Vet Immunol Immunopathol 121(3-4):370-7.
96. Zheng, K., P.G Board, X. Fei, et al. (2008) A novel selenium-containing glutathione transferase zetal-1, the activity of which surpasses the level of some native glutathione peroxidases. Int J Biochem Cell Biol.
97. Padhi, A. B. Verghese (2008) Detecting molecular adaptation at individual codons in the pattern recognition protein, lipopolysaccharide-and beta-1,3-glucan-binding protein of decapods. Fish Shellfish Immunol.
98. Jayaraj, S.S., R. Thiagarajan, M. Arumugam, et al. (2007) Isolation, purification and characterization of β 1,3-glucan binding protein from the plasma of marine mussel Perna viridis,. Fish and Shelfish Immunology
99. Luo, T., X. Zhang, Z. Shao, et al. (2003) PmAV, a novel gene involved in virus resistance of shrimp Penaeus monodon. FEBS Lett 551(1-3):53-7.
100. Luo, T., F. Li, K. Lei, et al. (2007) Genomic organization, promoter characterization and expression profiles of an antiviral gene PmAV from the shrimp Penaeus monodon. Mol Immunol 44(7):1516-23.
101. Sun, YD., L.D. Fu, Y.P. Jia, et al. (2008) A hepatopancreas-specific C-type lectin from the Chinese shrimp Fenneropenaeus chinensis exhibits antimicrobial activity. Mol Immunol 45(2):348-61.
102. Vasta, G.R., M. Quesenberry, H. Ahmed, et al. (1999) C-type lectins and galectins mediate innate and adaptive immune functions:their roles in the complement activation pathway. Dev Comp Immunol 23(4-5):401-20.
103. Argraves, W.S., L.M. Greene, M.A. Cooley, et al. (2003) Fibulins:physiological and disease perspectives. EMBO Rep 4(12):1127-31.
104. Clavero-Salas, A., R.R. Sotelo-Mundo, T. Gollas-Galvan, et al. (2007) Transcriptome analysis of gills from the white shrimp Litopenaeus vannamei infected with White Spot Syndrome Virus. Fish Shellfish Immunol 23(2):459-72.
105. Lee, S.Y., B.L. Lee K. Soderhall (2004) Processing of crayfish hemocyanin subunits into phenoloxidase. Biochem Biophys Res Commun 322(2):490-6.
106. Yang, J., Z.L. Wang, X.Q. Zhao, et al. (2008) Natural selection and adaptive evolution of leptin in the ochotona family driven by the cold environmental stress. PLoS One 3(1):e1472.
107. Yuen, B.S., P.C. Owens, M.E. Symonds, et al. (2004) Effects of leptin on fetal plasma adrenocorticotropic hormone and cortisol concentrations and the timing of parturition in the sheep. Biol Reprod 70(6):1650-7.
108. Paula-Lopes, F.F., M. Boelhauve, F.A. Habermann, et al. (2007) Leptin promotes meiotic progression and developmental capacity of bovine oocytes via cumulus cell-independent and-dependent mechanisms. Biol Reprod 76(3):532-41.
109. Paz-Filho, G.J., T. Babikian, R. Asarnow, et al. (2008) Leptin replacement improves cognitive development. PLoS One 3(8):e3098.
110. Kronfeld-Schor, N., J. Zhao, B.A. Silvia, et al. (2000) Steroid-dependent up-regulation of adipose leptin secretion in vitro during pregnancy in mice. Biol Reprod 63(1):274-80.
111. Henson, M.C. V.D. Castracane (2006) Leptin in pregnancy:an update. Biol Reprod 74(2):218-29.
112. Kratzsch, J., M. Hockel W. Kiess (2000) Leptin and pregnancy outcome. Curr Opin Obstet Gynecol 12(6):501-5.
113. Smith, G.D., L.M. Jackson D.L. Foster (2002) Leptin regulation of reproductive function and fertility. Theriogenology 57(1):73-86.
114. Magarinos, M.P., V. Sanchez-Margalet, M. Kotler, et al. (2007) Leptin promotes cell proliferation and survival of trophoblastic cells. Biol Reprod 76(2):203-10.
115. Morash, B.A., A. Imran, D. Wilkinson, et al. (2003) Leptin receptors are developmentally regulated in rat pituitary and hypothalamus. Mol Cell Endocrinol 210(1-2):1-8.
116. Perez-Perez, A., J. Maymo, J.L. Duenas, et al. (2008) Leptin prevents apoptosis of trophoblastic cells by activation of MAPK pathway. Arch Biochem Biophys 477(2):390-5.
117. Huang, Y., K. Ying, Y. Xie, et al. (2001) Cloning and characterization of a novel human leptin receptor overlapping transcript-like 1 gene (LEPROTLl). Biochim Biophys Acta 1517(2):327-31.
118. Rudnick, D.A., K. Hieb, K.F. Grimmer, et al. (2003) Patterns and processes of biological invasion:The Chinese mitten crab in San Francisco Bay. Basic and Applied Ecology 4(3):249-262.
119. Sui, L., M. Wille, Y. Cheng, et al. (2007) The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae. Aquaculture 273(1):139-150.
120. Wang, H.-Z., H.-J. Wang, X.-M. Liang, et al. (2006) Stocking models of Chinese mitten crab (Eriocheir japonica sinensis) in Yangtze lakes. Aquaculture 255(1-4):456-465.
121. Herborg, L.-M., M.G. Bentley, A.S. Clare, et al. (2006) Mating behaviour and chemical communication in the invasive Chinese mitten crab Eriocheir sinensis. Journal of Experimental Marine Biology and Ecology 329(1):1-10.
122. Li, X., Y. Lei, X. Gao, et al. (2007) Calcium carbonate supersaturation and precipitation in Chinese mitten crab (Eriocheir japonica sinensis) larval ponds in China:Mass mortality, crystal form analysis, and safety saturation index. Aquaculture 272(1-4):361-369.
123. Wen, X.B., L.Q. Chen, Z.L. Zhou, et al. (2002) Reproduction response of Chinese mitten-handed crab (Eriocheir sinensis) fed different sources of dietary lipid. Comparative Biochemistry and Physiology-Part A:Molecular & Integrative Physiology 131(3):675-681.
124. Kalinina, M.V., N.A. Vinnikova E.G. Semen'kova (2008) [Gonadogenesis and color characteristics of ovaries in Japanese mitten crab Eriocheir japonicus]. Ontogenez 39(1):58-65.
125. Wheelan, S.J., D.M. Church J.M. Ostell (2001) Spidey:a tool for mRNA-to-genomic alignments. Genome Res 11(11):1952-7.
126. Churbanov, A., M. Pauley, D. Quest, et al. (2005) A method of precise mRNA/DNA homology-based gene structure prediction. BMC Bioinformatics 6:261.
127. Assumpc-a-o, T.C.F., I.M.B. Francischetti, J.F. Andersen, et al. (2008) An insight into the sialome of the blood-sucking bug Triatoma infestans, a vector of Chagas' disease. Insect Biochemistry and Molecular Biology 38(2):213-232.
128. Kumar, S., K. Tamura M. Nei (2004) MEGA3:Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5(2):150-63.
129. Wang, Y, K.K. Kuropatwinski, D.W. White, et al. (1997) Leptin receptor action in hepatic cells. J Biol Chem 272(26):16216-23.
130. Cheng, S.P., C.W. Chi, C.Y. Tzen, et al. (2009) Clinicopathologic significance of leptin and leptin receptor expressions in papillary thyroid carcinoma. Surgery.
131. Zhao, J., L. Song, C. Li, et al. (2007) Molecular cloning, expression of a big defensin gene from bay scallop Argopecten irradians and the antimicrobial activity of its recombinant protein. Mol Immunol 44(4):360-8.
132. Fang, J.J. G.F. Qiu (2009) Molecular cloning of cyclin B transcript with an unusually long 3' untranslation region and its expression analysis during oogenesis in the Chinese mitten crab, Eriocheir sinensis. Mol Biol Rep 36(6):1521-9.
133. Mollineau, W.M., A.O. Adogwa G.W. Garcia (2009) The gross and micro anatomy of the accessory sex glands of the male agouti (Dasyprocta leporina). Anat Histol Embryol 38(3):204-7.
134. Trinadha Babu, B., K. Shyamasundari K. Hanumantha Rao (1989) Studies of the neurohaemal organ (sinus gland) of the portunid crab Portunus sanguinolentus (Herbst) (Brachyura).1. Morphology and histology in relation to moulting. Folia Morphol (Praha) 37(4):388-93.
135. Zarubin, T.P., E.S. Chang D.L. Mykles (2009) Expression of recombinant eyestalk crustacean hyperglycemic hormone from the tropical land crab, Gecarcinus lateralis, that inhibits Y-organ ecdysteroidogenesis in vitro. Mol Biol Rep 36(6):1231-7.
136. Normant, M., M. Chrobak A. Szaniawska (2002) Energy value and chemical composition (CHN) of the Chinese mitten crab Eriocheir sinensis (Decapoda:Grapsidae) from the Baltic Sea. Thermochimica Acta 394(1-2):233-237.
137. Yamaguchi, S., F.T. Celino, A. Ito, et al. (2008) Effects of arsenic on gonadal development in freshwater crab, Somanniathelphusa pax, in Vietnam and Geothelphusa dehaani in Japan. Ecotoxicology 17(8):772-80.
138. Henault, M.A., G.J. Killian, J.F. Kavanaugh, et al. (1995) Effect of accessory sex gland fluid from bulls of differing fertilities on the ability of cauda epididymal sperm to penetrate zona-free bovine oocytes. Biol Reprod 52(2):390-7.
139. Abd el-Hamid, N.F. (1988) Size composition, sex ratio, size at maturity, mating and egg-laying in the crab Portunus pelagicus (Linnaeus). Folia Morphol (Praha) 36(2):200-10.
140. Le Bizec, B., J.-P. Antignac, F. Monteau, et al. (2002) Ecdysteroids:one potential new anabolic family in breeding animals. Analytica Chimica Acta 473(1-2):89-97.
141. Margam, V.M., D.B. Gelman S.R. Palli (2006) Ecdysteroid titers and developmental expression of ecdysteroid-regulated genes during metamorphosis of the yellow fever mosquito, Aedes aegypti (Diptera:Culicidae). Journal of Insect Physiology 52(6):558-568.
142. Burtis, K.C., C.S. Thummel, C.W. Jones, et al. (1990) The Drosophila 74EF early puff contains E74, a complex ecdysone-inducible gene that encodes two ets-related proteins. Cell 61(1):85-99.
143. Sekimoto, T., M. Iwami S. Sakurai (2007) 20-Hydroxyecdysone regulation of two isoforms of the Ets transcription factor E74 gene in programmed cell death in the silkworm anterior silk gland. Insect Mol Biol 16(5):581-90.
144. Hepperle, C. K. Hartfelder (2001) Differentially expressed regulatory genes in honey bee caste development. Naturwissenschaften 88(3):113-6.
145. Pongs, O. (1988) Ecdysteroid-regulated gene expression in Drosophila melanogaster. Eur J Biochem 175(2):199-204.
146. Paul, R.K., H. Takeuchi, Y. Matsuo, et al. (2005) Gene expression of ecdysteroid-regulated gene E74 of the honeybee in ovary and brain. Insect Mol Biol 14(1):9-15.
147. Mu, X.Y. G.A. Leblanc (2004) Cross communication between signaling pathways:Juvenoid hormones modulate ecdysteroid activity in a crustacean. Journal of Experimental Zoology Part a-Comparative Experimental Biology 301A(10):793-801.
148. Matsuoka, T. H. Fujiwara (2000) Expression of ecdysteroid-regulated genes is reduced specifically in the wing discs of the wing-deficient mutant (fl) of Bombyx mori. Development Genes and Evolution 210(3):120-128.
149. Arnold, K., L. Bordoli, J. Kopp, et al. (2006) The SWISS-MODEL workspace:a web-based environment for protein structure homology modelling. Bioinformatics 22(2):195-201.
150. Schwede, T., J. Kopp, N. Guex, et al. (2003) SWISS-MODEL:An automated protein homology-modeling server. Nucleic Acids Res 31(13):3381-5.
151. Guex, N. M.C. Peitsch (1997) SWISS-MODEL and the Swiss-PdbViewer:an environment for comparative protein modeling. Electrophoresis 18(15):2714-23.
152. Liou, H.L., S.S. Dixit, S. Xu, et al. (2006) NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. J Biol Chem 281 (48):36710-23.
153. Maroy, P., R. Dennis, C. Beckers, et al. (1978) Demonstration of an ecdysteroid receptor in a cultured cell line of Drosophila melanogaster. Proc Natl Acad Sci U S A 75(12):6035-8.
154. Yund, M.A., D.S. King J.W. Fristrom (1978) Ecdysteroid receptors in imaginal discs of Drosophila melanogaster. Proc Natl Acad Sci U S A 75(12):6039-43.
155. Lan, Q., Z. Wu L.M. Riddiford (1997) Regulation of the ecdysone receptor, USP, E75 and MHR3 mRNAs by 20-hydroxyecdysone in the GVI cell line of the tobacco hornworm, Manduca sexta. Insect Mol Biol 6(1):3-10.
156. Barker, G.C. H.H. Rees (1990) Ecdysteroids in nematodes. Parasitol Today 6(12):384-7.
157. Mathias, J.D., Y. Ran, J.D. Carter, et al. (2009) Interactions of the GM2 activator protein with phosphatidylcholine bilayers:a site-directed spin-labeling power saturation study. Biophys J 97(5):1436-44.
158. Jelinek, D., S.M. Patrick, K.N. Kitt, et al. (2009) Physiological and coordinate downregulation of the NPC1 and NPC2 genes are associated with the sequestration of LDL-derived cholesterol within endocytic compartments. J Cell Biochem 108(5):1102-16.
159. Record, E., S. Moukha M. Asther (1999) Characterization and expression of the cDNA encoding a new kind of phospholipid transfer protein, the phosphatidylglycerol/phosphatidylinositol transfer protein from Aspergillus oryzae:evidence of a putative membrane targeted phospholipid transfer protein in fungi. Biochim Biophys Acta 1444(2):276-82.
160. Ichikawa, S., T. Takai, T. Yashiki, et al. (2009) Lipopolysaccharide binding of the mite allergen Der f 2. Genes Cells 14(9):1055-65.
161. Park, S.Y., J.H. Cho, D.Y. Oh, et al. (2009) House dust mite allergen Der f 2-induced phospholipase Dl activation is critical for the production of interleukin-13 through activating transcription factor-2 activation in human bronchial epithelial cells. J Biol Chem 284(30):20099-110.
162. Xie, B., W. Wang D.J. Mahuran (1992) A Cys138-to-Arg substitution in the GM2 activator protein is associated with the AB variant form of GM2 gangliosidosis. Am J Hum Genet 50(5):1046-52.
163. Elleder, M., H. Houstkova, J. Zeman, et al. (2001) Pulmonary storage with emphysema as a sign of Niemann-Pick type C2 disease (second complementation group). Report of a case. Virchows Arch 439(2):206-11.
164. Xu, S., B. Benoff, H.L. Liou, et al. (2007) Structural basis of sterol binding by NPC2, a lysosomal protein deficient in Niemann-Pick type C2 disease. J Biol Chem 282(32):23525-31.
165. Babalola, J.O., M. Wendeler, B. Breiden, et al. (2007) Development of an assay for the intermembrane transfer of cholesterol by Niemann-Pick C2 protein. Biol Chem 388(6):617-26.
166. Busso, D., MJ. Onate-Alvarado, E. Balboa, et al. (2009) Female infertility due to anovulation and defective steroidogenesis in NPC2 deficient mice. Mol Cell Endocrinol.
167. Friedland, N., H.L. Liou, P. Lobel, et al. (2003) Structure of a cholesterol-binding protein deficient in Niemann-Pick type C2 disease. Proc Natl Acad Sci U S A 100(5):2512-7.
168. Kataoka, H., H. Nagasawa, A. Isogai, et al. (1991) Prothoracicotropic hormone of the silkworm, Bombyx mori:amino acid sequence and dimeric structure. Agric Biol Chem 55(1):73-86.
169. Marchal, E., H.P. Vandersmissen, L. Badisco, et al. (2009) Control of ecdysteroidogenesis in prothoracic glands of insects:a review. Peptides 31(3):506-19.
170. Covi, J.A., B.D. Bader, E.S. Chang, et al. Molt cycle regulation of protein synthesis in skeletal muscle of the blackback land crab, Gecarcinus lateralis, and the differential expression of a myostatin-like factor during atrophy induced by molting or unweighting. J Exp Biol 213(Pt1):172-83.
171. Mykles, D.L. D.M. Skinner (1982) Molt cycle-associated changes in calcium-dependent proteinase activity that degrades actin and myosin in crustacean muscle. Dev Biol 92(2):386-97.
172. Zmora, N., J. Trant, Y. Zohar, et al. (2009) Mo It-inhibiting hormone stimulates vitellogenesis at advanced ovarian developmental stages in the female blue crab, Callinectes sapidus 1:an ovarian stage dependent involvement. Saline Systems 5:7.
173. Reddy, P.S., A. Bhagylakshmi R. Ramamurthi (1985) Molt-inhibition in the crab Oziotelphusa senex senex following exposure to malathion and methyl parathion. Bull Environ Contam Toxicol 35(1):92-7.
174. Han, D.W., N. Patel R. Douglas Watson (2006) Regulation of protein synthesis in Y-organs of the blue crab (Callinectes sapidus):involvement of cyclic AMP. J Exp Zool A Comp Exp Biol 305(4):328-34.
175. Spindler, K.D., D. Adelung C. Tchernigovtzeff (1974) A comparison of the methods of molt staging according to Drach and to Adelung in the common shore crab, Carcinus maenas. Z Naturforsch C 29(11-12):754-6.
176. Nakatsuji, T., C.Y. Lee R.D. Watson (2009) Crustacean molt-inhibiting hormone:structure, function, and cellular mode of action. Comp Biochem Physiol A Mol Integr Physiol 152(2):139-48.
177. Subramoniam, T. (2000) Crustacean ecdysteriods in reproduction and embryogenesis. Comp Biochem Physiol C Toxicol Pharmacol 125(2):135-56.
178. Meszaros, M. D.B. Morton (1997) Up-and downregulation of esr20, an ecdysteroid-regulated gene expressed in the tracheae of Manduca sexta. Arch Insect Biochem Physiol 34(2):159-74.
179. Paul, R.K., H. Takeuchi T. Kubo (2006) Expression of two ecdysteroid-regulated genes, Broad-Complex and E75, in the brain and ovary of the honeybee (Apis mellifera L.). Zoolog Sci 23(12):1085-92.
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