乌鳢感染A.veronii差异基因的筛选及代表性基因在体内表达规律研究
|
摘要
乌鳢作为食补与药补于一身的名贵鱼类,在我国深受广大消费者的喜爱。近年来,随着乌鳢养殖面积的增加、集约化养殖的发展,其病害也逐年增多,尤其是细菌性疾病日趋严重,已成为制约乌鳢养殖业健康可持续发展的重大障碍。目前,有关乌鳢细菌性疾病的治疗多采用投喂抗生素的方法,但此方法易导致乌鳢体内药物残留以及细菌耐药性增加。研发新型无毒渔药与利用免疫相关基因进行抗病品系选育是乌鳢养殖业健康可持续发展的趋势,因此,必须对乌鳢抗病免疫相关基因及免疫机理进行深入研究。
本研究应用SSH技术,成功构建维氏气单胞菌感染后的乌鳢头肾SSH cDNA文库,并经筛选、测序,获得有效序列202条,其中,73个序列未搜到同源性,其余129条序列按可能功能共分为6大类,免疫相关基因序列25条,占总数的13%。
在乌鳢头肾SSH cDNA文库的基础上,从中选出疑似Cu/Zn-SOD、Rac2、MPO的基因片段,并利用RACE技术获得了三种免疫相关基因的全长cDNA序列,采用生物信息学方法对其理化性质、蛋白结构及氨基酸同源性进行了分析；用实时荧光定量PCR技术分析了免疫相关基因在健康乌鳢的不同组织中表达分布以及感染维氏气单胞菌后的表达变化。结果显示：
Cu/Zn-SOD基因全长cDNA序列810bp,其ORF465bp、编码154个氨基酸；多重序列比较分析,Cu/Zn-SOD氨基酸序列与莫桑比克罗非鱼同源性为95%；实时荧光定量PCR分析表达显示：Cu/Zn-SOD基因在健康乌鳢的头肾、肾、鳃、心、脾、肝、肌肉、肠等八个组织中均有表达,肝脏中的表达量最高；感染维氏气单胞菌后,其在不同组织中的表达量变化大致呈现：先上升后下降并维持一段时间,后又有上升的趋势。
Rac2基因全长cDNA序列1179bp,其ORF579bp、编码192个氨基酸；通过氨基酸序列比较分析,各物种Rac2高度同源；实时荧光定量PCR分析表达显示：Rac2基因在健康乌鳢的头肾、肾、鳃、心、脾、肝、肌肉等七个组织中均有表达,不在肠中表达,其中,头肾表达量最高；感染维氏气单胞菌后,Rac2在乌鳢7个组织中的表达量,随感染时间不同而发生变化,但均在感染后的36h时,表达量上调,基本超过0h时的表达量。
MPO基因全长cDNA序列3181bp,其ORF2301bp、编码766个氨基酸；通过氨基酸序列比较分析,各物种MPO蛋白存在歧化；实时荧光定量PCR分析表达显示：MPO基因在健康乌鳢的头肾、肾、鳃、心、脾、肝、肌肉等七个组织中均有表达,在肠中不表达,其中,头肾表达量最高；感染维氏气单胞菌后,MPO在大多数组织中低水平表达；所有组织中一般在感染后的36h时,表达量有上升的趋势。
本研究成功的构建了乌鳢头肾SSH cDNA文库,并获得了大量与乌鳢抗菌感染相关的基因片段,在此基础上,应用RACE技术获得三种免疫相关基因的全长,利用生物信息学对其进行了较为系统的分析,同时探讨了这三种基因的表达模式。研究成果为进一步了解并研究乌鳢抗菌感染的分子免疫机制奠定基础,同时为抗病品种的选育及病害防治提供了基础理论数据。
Channa argus, one of valuable fishes which is loved by the customers, is been food and medicine. In recent years, more kinds of diseases occur with the incremental farming area and the intensive aquaculture, especially the bacterial disease. This has become a significant barrier to the aquaculture development of Channa argus. Currently, to feed antibiotics is the general method to cure the bacterial disease. However, the drug residue and resistance is increasing. To develop the new no-toxic drug and disease resistance breeding is the tendency of the sustainable development of Channa argus aquaculture. Hence, it requires further study to antibacterial immunity-related genes and the immunologic mechanism of Channa argus.
In this research, the head-kidney SSH cDNA library from Channa argus infected with Aeromonas veronii was constructed using Suppression Subtractive Hybridization. And by screening and BLASTn,202expressed sequence tags(ESTs) were acquired.73ESTs were unknown sequence, and the other129ESTs were grouped under6clusters based on their function. The immunity-related genes consist of25ESTs and represented13%of the whole ESTs.
From the head-kidney SSH library, the gene segment of Cu/Zn-SOD, Rac2, and MPO were picked, and the full length were acquired with RACE. By the bioinformatics, the chemicophysical properties, protein structure and amino acid homology were analyzed. The expression and distribution of the three genes in health Channa argus and the expression change after challenge with Aeromonas veronii were detected by real-time quantitative PCR (RT-PCR).
These results showed that the full length of Cu/Zn-SOD gene contained810bases, whose ORF contained465bases and encoded154amino acids. The multiple sequence alignment showed the amino acid sequence of Cu/Zn-SOD shared95%homologies with Tilapia Mossambica. The RT-PCR results showed that the SOD was expressed in head-kidney, kidney, gill, heart, spleen, liver, muscle, and intestines, and the relative expression quality of liver was the highest. After challenge with Aeromonas veronii, the average trend of expression quality was after rising to decline and maintained for a period of time, and then rose.
The full length of Rac2gene contained1179bases, whose ORF contained579bases and encoded192amino acids. The amino acid sequence showed that the Rac2from most species all shared high homologies. The RT-PCR results showed that the Rac2was expressed in head-kidney, kidney, gill, heart, spleen, liver, and muscle, but not in intestines, and the relative expression quality of head-kidney was the highest. After challenge with Aeromonas veronii, the expression quality changed with time and all of them rose at36hour post-infection and exceeded it at0hr P.I.
The full length of MPO gene contained3181bases, whose ORF contained2301bases and encoded766amino acids. The amino acid sequence showed that the MPO from most species was different from one another. The RT-PCR results showed that the Rac2was expressed in head-kidney, kidney, gill, heart, spleen, liver, and muscle, but not in intestines, and the relative expression quality of head-kidney was the highest. After challenge with Aeromonas veronii, the expression quality was low in most tissue and all rose at36hr P.I..
In this work, we successfully constructed the head-kidney SSH cDNA library from Channa argus, and a number of ESTs related to antibacterial immunity was acquired. The full length of three genes was amplified and systematic analyzed, and then their patterns of expression were explored. These results can lay the foundation for further study about the molecular mechanism of immunity and the basic theoretical data for resistance breeding and disease control.
本研究应用SSH技术,成功构建维氏气单胞菌感染后的乌鳢头肾SSH cDNA文库,并经筛选、测序,获得有效序列202条,其中,73个序列未搜到同源性,其余129条序列按可能功能共分为6大类,免疫相关基因序列25条,占总数的13%。
在乌鳢头肾SSH cDNA文库的基础上,从中选出疑似Cu/Zn-SOD、Rac2、MPO的基因片段,并利用RACE技术获得了三种免疫相关基因的全长cDNA序列,采用生物信息学方法对其理化性质、蛋白结构及氨基酸同源性进行了分析；用实时荧光定量PCR技术分析了免疫相关基因在健康乌鳢的不同组织中表达分布以及感染维氏气单胞菌后的表达变化。结果显示：
Cu/Zn-SOD基因全长cDNA序列810bp,其ORF465bp、编码154个氨基酸；多重序列比较分析,Cu/Zn-SOD氨基酸序列与莫桑比克罗非鱼同源性为95%；实时荧光定量PCR分析表达显示：Cu/Zn-SOD基因在健康乌鳢的头肾、肾、鳃、心、脾、肝、肌肉、肠等八个组织中均有表达,肝脏中的表达量最高；感染维氏气单胞菌后,其在不同组织中的表达量变化大致呈现：先上升后下降并维持一段时间,后又有上升的趋势。
Rac2基因全长cDNA序列1179bp,其ORF579bp、编码192个氨基酸；通过氨基酸序列比较分析,各物种Rac2高度同源；实时荧光定量PCR分析表达显示：Rac2基因在健康乌鳢的头肾、肾、鳃、心、脾、肝、肌肉等七个组织中均有表达,不在肠中表达,其中,头肾表达量最高；感染维氏气单胞菌后,Rac2在乌鳢7个组织中的表达量,随感染时间不同而发生变化,但均在感染后的36h时,表达量上调,基本超过0h时的表达量。
MPO基因全长cDNA序列3181bp,其ORF2301bp、编码766个氨基酸；通过氨基酸序列比较分析,各物种MPO蛋白存在歧化；实时荧光定量PCR分析表达显示：MPO基因在健康乌鳢的头肾、肾、鳃、心、脾、肝、肌肉等七个组织中均有表达,在肠中不表达,其中,头肾表达量最高；感染维氏气单胞菌后,MPO在大多数组织中低水平表达；所有组织中一般在感染后的36h时,表达量有上升的趋势。
本研究成功的构建了乌鳢头肾SSH cDNA文库,并获得了大量与乌鳢抗菌感染相关的基因片段,在此基础上,应用RACE技术获得三种免疫相关基因的全长,利用生物信息学对其进行了较为系统的分析,同时探讨了这三种基因的表达模式。研究成果为进一步了解并研究乌鳢抗菌感染的分子免疫机制奠定基础,同时为抗病品种的选育及病害防治提供了基础理论数据。
Channa argus, one of valuable fishes which is loved by the customers, is been food and medicine. In recent years, more kinds of diseases occur with the incremental farming area and the intensive aquaculture, especially the bacterial disease. This has become a significant barrier to the aquaculture development of Channa argus. Currently, to feed antibiotics is the general method to cure the bacterial disease. However, the drug residue and resistance is increasing. To develop the new no-toxic drug and disease resistance breeding is the tendency of the sustainable development of Channa argus aquaculture. Hence, it requires further study to antibacterial immunity-related genes and the immunologic mechanism of Channa argus.
In this research, the head-kidney SSH cDNA library from Channa argus infected with Aeromonas veronii was constructed using Suppression Subtractive Hybridization. And by screening and BLASTn,202expressed sequence tags(ESTs) were acquired.73ESTs were unknown sequence, and the other129ESTs were grouped under6clusters based on their function. The immunity-related genes consist of25ESTs and represented13%of the whole ESTs.
From the head-kidney SSH library, the gene segment of Cu/Zn-SOD, Rac2, and MPO were picked, and the full length were acquired with RACE. By the bioinformatics, the chemicophysical properties, protein structure and amino acid homology were analyzed. The expression and distribution of the three genes in health Channa argus and the expression change after challenge with Aeromonas veronii were detected by real-time quantitative PCR (RT-PCR).
These results showed that the full length of Cu/Zn-SOD gene contained810bases, whose ORF contained465bases and encoded154amino acids. The multiple sequence alignment showed the amino acid sequence of Cu/Zn-SOD shared95%homologies with Tilapia Mossambica. The RT-PCR results showed that the SOD was expressed in head-kidney, kidney, gill, heart, spleen, liver, muscle, and intestines, and the relative expression quality of liver was the highest. After challenge with Aeromonas veronii, the average trend of expression quality was after rising to decline and maintained for a period of time, and then rose.
The full length of Rac2gene contained1179bases, whose ORF contained579bases and encoded192amino acids. The amino acid sequence showed that the Rac2from most species all shared high homologies. The RT-PCR results showed that the Rac2was expressed in head-kidney, kidney, gill, heart, spleen, liver, and muscle, but not in intestines, and the relative expression quality of head-kidney was the highest. After challenge with Aeromonas veronii, the expression quality changed with time and all of them rose at36hour post-infection and exceeded it at0hr P.I.
The full length of MPO gene contained3181bases, whose ORF contained2301bases and encoded766amino acids. The amino acid sequence showed that the MPO from most species was different from one another. The RT-PCR results showed that the Rac2was expressed in head-kidney, kidney, gill, heart, spleen, liver, and muscle, but not in intestines, and the relative expression quality of head-kidney was the highest. After challenge with Aeromonas veronii, the expression quality was low in most tissue and all rose at36hr P.I..
In this work, we successfully constructed the head-kidney SSH cDNA library from Channa argus, and a number of ESTs related to antibacterial immunity was acquired. The full length of three genes was amplified and systematic analyzed, and then their patterns of expression were explored. These results can lay the foundation for further study about the molecular mechanism of immunity and the basic theoretical data for resistance breeding and disease control.
引文
[1]刘苏,朱新平,陈昆慈,等.斑鳢、乌鳢及其杂种形态差异分析[J].华中农业大学学报,2011,30(4)：488-493.
[2]王广军.乌鳢的生物学特性及繁殖技术[J].淡水渔业,2000,30(6)：10-11.
[3]聂国兴,傅艳茹,张浩,等.乌鳢肌肉营养成分分析[J].淡水渔业,2002,32(2)：46-47.
[4]吴同垒,单晓枫,王桂芹,等.乌鳢主要病害研究进展[J].黑龙江畜牧兽医,2011,9：29-31.
[5]潘立新,郑善坚.乌鳢暴发性出血病的诊断和治疗[J].淡水渔业,2004，34(4)：49-50.
[6]曹铮,沈锦玉,谢荣林,等.乌鳢结节病病原-诺卡氏菌的分离与鉴定[J].水生生物学报,2009,33(2)：338-340.
[7]黄江峰,饶毅,金友萍,等.乌鳢溃疡综合症流行病规律调查[J].江西水产科技,2008,4：37-39.
[8]张海宾.乌鳢出血性败血症病原研究[J].水利渔业,2005,25(2)：81-82.
[9]舒新华,金燮理,肖克宇,等.乌鳢腹水病病原的分离和鉴定[J].湖南农业大学学报,1998,24(4)：286-290.
[10]曾慷,舒新华,金燮理.乌鳢腐皮病致病细菌的研究[J].湖南农业大学学报,1999,25(5)：396-399.
[11]舒新华,曾慷,金燮理,等.乌鳢腐皮病病理形态学研究[J].淡水渔业,1999,29(8)：8-10.
[12]袁思平,王国良,金珊.养殖鱼类致病诺卡氏菌研究进展[J].微生物通报,2006,33(2)：137-141.
[13]王家沛.乌鳢诺卡氏菌病的诊治[J].现代农业科技,2010(13)：387-390.
[14]吕伟清,孙伟杰,谢荣林.乌鳢诺卡氏菌的防治[J].科学养鱼,2008(1)：57.
[15]王国良,徐益军,金珊,等.养殖乌鳢诺卡氏菌病及其病原研究[J].水生生物学报,2009,33(2)：277-283.
[16]石存斌,潘厚军,常藕琴,等.养殖乌鳢结节病的病原分析[J].安徽农业科学,2009,37(16)：7384-7386.
[17]胡银亨.乌鳢车轮虫研究[J].泸州职业技术学院学报,2007,4：34-37.
[18]Mauel M J, Miller D L. Piscirickettsiosis and piscirickettsiosis-like infections in fish:a review[J].Vet Microbiol,2002,87:279-289.
[19]贾伟章,孙晓凤,郭琼林.养殖乌鳢类立克次体分离纯化的初步研究[J].水生生物学学报,2004,28(4)：448-449.
[20]Guo Q L, Jia W Z, Han X P, et al. Rickettsia-like organism infection in a freshwater cultured fish Ophiocephalus argus C. in China [J]. Prog Nat Sci.2004,14:417-422.
[21]单晓枫,郭伟生,张洪波,等.鱼类体液中的几种抗菌因子研究进展[J].河南农业科学,2010,5：138-142.
[22]Claire M H, Lambris J D. The complement system in teleosts [J].Fish&Shellfish Immunology,2002, 12:399-420.
[23]Kuroda N, Naruse K, Shima A, et al. Molecular cloning and linkage analysis of comp lement C3 and C4 genes of the Japanese medakafish [J]. Immunogenetics.2000,51:117-128.
[24]NakaoM, Mutsuro J, Obo R, et al. Molecular cloning and protein analysis of divergent forms of the complement component C3 from a bony fish, the common carp (Cyprinus carpio):presence of variants lacking the catalytic histidine[J]. Eur J Immunol,2000,30:858-866.
[25]Smith S H, Jensen J A. The second component (C2n) of the nurse shark complement system:purification, physico-chemical characterization and functional comparison with guinea pig C4 [J]. Dev Comp Immunol,1986,10:191-206.
[26]Terado T, Okamura K, Ohta Y, et al. Molecular cloning of C4 gene and identification of the classIII complement region in the shark MHC [J]. J Immunol,2003,171:2461-2466.
[27]Shin D H, Webb B, NakaoM, et al. Molecular cloning, structural analysis and expression of complement component B f/C2 genes in the nurse shark, Ginglym ostom a cirratum[J]. Dev Comp Immunol,2007,, 31:1168-1182.
[28]Wang T, Secombes C J. Complete sequencing and expression of three complement components, Clr, C4 and Cl inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss [J]. Immunogenetics,2003,55:615-628.
[29]Ellingsen T, Strand C, Monsen E, et al. The ontogeny of complement component C3 in the spotted wolfish (Anarhichasminor olafsen) [J]. Fish Shellfish Immunol,2005,18:351-358.
[30]Lange S, Slavko H B, Alister W D, et al. Complement component C3 transcription in Atlantic halibut (Hippoglossus hippoglossus L.) larvae [J]. Fish Shellfish Immunol,2006,20:285-294.
[31]Marie Lovoll, Terje Kilvik, HaniBoshra, et al. Maternal transfer of complement components C3-1, C3-3, C3-4, C4, C5, C7, Bf and Df to off spring in rainbow trout (Oncorhynchus mykiss) [J].Immunogenetics, 2006,58:168-179.
[32]Fujita T. Evolution of lecfin-complement pathway and its role in innate immunity [J]. Nature,2002,2: 347-353.
[33]Boshra H, Li J, Sunyer J O. Recent advances on the complement system of teleost fish [J]. Fish Shellfish Immunol,2006,20:239-262.
[34]Boshra H, Gelman A E, Sunyer J O. Structural and functional characterization of complement C4 and Cls-like molecules in teleost fish:insights into the evolution of classical and alternative pathways [J]. J Immunol,2004,173:349-359.
[35]Endo Y, Takahashi M, Fujita T. Lectin complement system and pattern recognition [J]. Immunolobiology,2006,211:283-293.
[36]Boshra H, Peters R, Li J, et al. Production of recombinant C5a from rainbow trout (Oncorhynchus mykiss):role in leucocyte chemotaxis and respiratory burst [J]. Fish Shellfish Immunol,2004,17: 293-303.
[37]Barnes A C, Ellis A E. Role of capsule in serotypic differences and complement fixation by lactococcus garvieae[J]. Fish Shellfish Immunol,2004,16:207-214.
[38]Fernandez-Trujillo M A, Bejar J, Gallardo J B, et al. Molecular cloning and characterization of C-type lysozyme from Senegalese sole (Solea senegalensis) [J]. Aquaculture,2007,272 (Supp 11):255.
[39]Fast M D, Sims D E, Burka J F, et al. Skin morphology and humoral non-specific defence parameters of mucus and plasma in rainbow trout, coho and Atlantic salmon[J]. Comp Biochem Physiol A Mol Integr Physiol,2002,132(3):645-657.
[40]刘红柏,魏巍.史氏鲟及小体鲟不同组织中溶菌酶水平的比较[J].水产学杂志,2007,20(1)：57-60.
[41]Subbotkina TA, Subbotkin M F. Lysozyme content in organs and blood serum in various species in the Volga River [J]. J. Evolutionary Biochemistry and Physiology,2003,39 (5):537-546.
[42]Hikima J, Minagawa S, Hirono I, et al. Molecular cloning, expression and evolution of the Japnanese flounder goose-type lysozyme gene, and the lytic activity of its recombinant protein[J]. Biochimica et Biophysica Acta,2001,1520:35-44.
[43]Mitra A, Foster-Frey J, Rexroad C E, et al. Molecular characterization of lysozyme type Ⅱ gene in rainbow trout (Oncorhynchus mykiss):evidence of gene duplication[J].Anim Biotechnol,2003,14 (1):7-12.
[44]Yin Z X, He J G, Deng W X, et al. Molecular cloning, expression of orange-spotted grouper goose-type lysozyme cDNA, and lytic activity of its recombinant protein[J]. Dis Aquat Organ,2003,55 (2):117-123.
[45]纪伟,张培军.转“全鱼”溶菌酶基因大菱鲆的研究[J].海洋科学,2004,28(5)：8-14.
[46]Cho Y S, Lee S Y, Kim K Y, et al. Gene structure and expression of metallot hionein during metal exposures in Hemibarbus mylodon [J]. Ecotoxicol Environ Safety,2008,71(1):125-137.
[47]Schmidt C, Beyersmann D. Transient peaks in zinc and metalliothionein levels during differentiation of 3T3L1 cells [J].Arch Biochem Biophys,1999,364:91-98.
[48]Petrosillo G, Di Venosa N, Ruggiero F M, et al. Mitochondrial dysfunction associated wit h cardiac ischemia/reperfusion can be attenuated by oxygen tension control. Role of oxygenfree radicals and cardiolipin[J]. Biochim Biophy Acta Bioenerget,2005,1710(2-3):78-86.
[49]Haq F, Mahoney M, Koropatnick J. Signaling events for metallothionein induction [J]. Mutat Res,2003, 533:211-226.
[50]Bonhan k M, Zafarullahl, Gedamu. The rainbow trout metallothioneins molecular cloning and characterization of two distinct cDNA sequences [J]. DNA,1987, (6):519-528.
[51]赵晓祥,张淑梅,李晶,等.虹鳟鱼金属硫蛋白启动子的体外扩增、克隆及其序列分析[J].生物技术,2000,10(1)：1-3.
[52]冯仁勇,周小秋.鱼类金属硫蛋白基因的表达与调控[J].水产科学,2005,24(9)：52-54.
[53]龙华,曾勇.鱼血清转铁蛋白对嗜水气单胞菌的抗性研究[J].湖北农学院学报,2004,24(2)：119-123.
[54]FerixenetN, Remacha A, Berlanga E, et al. Serum soluble transferrin receptor concentrations are increased in central obesity. Results from a screening programme for hereditary hemochromatosis in men with hyperferritinemia[J]. Clinica Chimica Acta,2009,400:111-116.
[55]Park I, Schaeffer E, Sidoli A, et al. Organization of the human transferrin gene:Direct evidence that it originated by gene duplication[J]. Proc Natl Acad Sci U S A,1985,82:3149.
[56]Bartfeld N S, Law J H. Isolation and molecular cloning of transferring from the tobacco hornworm, Manduca Sexta[J].J Biol Chem,1990,265:21684-21691.
[57]Hirono I, Uchiyama T, Aoki T. Cloning, nucleotide sequence analysis, and characterization of cDNA for medaka (Oryzias latipes) transferring [J]. J Mar Biotechnol,1995,2:193-198.
[58]Lee J Y, Tange N, Yamashita H, et al. Cloning and characterization of transferrin cDNA from coho salmon (Oncorhynchus kisutch) [J]. Fish Pathology,1995,30(4):271-277.
[59]龙华.鲫血清转铁蛋白cDNA克隆及其序列分析[J].中山大学学报(自然科学版),2003,42(增刊)：231-234.
[60]李明云,陈炯,张祖兴,等.大黄鱼(Pseudosciaena crocea)转铁蛋白(Tf)基因克隆及序列分析[J].海洋与湖沼,2007,38(6)：563-567.
[61]林亚秋,郑玉才,吉红,等.草鱼转铁蛋白受体cDNA序列克隆及其组织表达差异[J].湖北农业科学,2009,48(6)：1289-1292.
[62]Fock W L, Chen C L, Lam TJ, et al. Roles of an endogenous serum lectin in immune protection of blue gourami, Trichogaster trichopteurs (Pallus) against Aeromonas hydrophila [J]. Fish Shellfish Immunol, 2001,11:245-252.
[63]Ottinger C A, Johnson S C, Ewart KV, et al. Enhancement of anti-Aeromonas salmonicida activity in Atlantic salmon (Salmo salar) macrophages by a mannose binding lectin [J].Comp Biochem Physiol C, 1999,123:53-59.
[64]Stratton L, WuS, Richards R C, et al. Oligomerisation and carbohydrate binding in an Atlantic salmon serum C-type lectin consistent with non-self recognition[J]. Fish Shellfish Immunol,2004,17:315-323.
[65]Okamoto M, Tsutsui S, Tasumi S, et al. Tandem repeat L-rhamnosebinding lectin from the skin mucus of ponyfish,Leiognathus nuchalis[J]. Biochem Bioph Res Co,2005,333:463-469.
[66]Jungw K, Park P J, Kim S K. Purification and characterization of a new lectin from the hard roe of skipjack tuna, Katsuwonus pelam is [J]. Int J Biochem Cell B,2003,35:255-265.
[67]吴超,陆承平.二龄草鱼脾脏、肝脏组织高表达甘露糖结合凝集素mRNA[J]动物学报,2004,50(1)：137-140
[68]Vitved L, Holmskov U, Koch C, et al. The homologue of mannose-binding lectin in the carp family Cyprinidae is expressed at high level in spleen, and deduced primary structure predicts affinity for galactose[J]. Immune-genetics,2000,51:955-964.
[69]吴超,陆承平.草鱼等8种淡水鱼血清凝集素作用的比较[J].南京农业大学学报,2004,27(2)：79-82.
[70]张黎,刘莉,袁媛,等.中华倒刺(?)血清凝集素的研究[J].南方水产,2006,2(6)：39-44.
[71]盘赛昆,顾小红,汤坚,等.鳙鳃凝集素的稳定性及细胞凝集作用[J].安徽农业科学,2009,37(26)：12589-12591.
[72]Huang P, Chen J, Kuo C.Three different hepcidins from tilapia,oreochromis mossambicus:Analysis of their expressions and biological functions[J]. Molecular Immunolgy,2007,44(8):1922-1934.
[73]Cuesta A, Meseguer J, Esteban M. The antimicrobial peptide hepcidin exerts an important role in the innate immunity against bacteria in the bony fish gilthead seabream [J]. Mol Immunol,2008, 45: 2333-2342.
[74]Wang KJ, Cai J J, Cai L, et al. Cloning and expression of a hepcidin gene from a marine fish (Pseudosciaena crocea) and the antimicrobial activity of it s synthetic peptide[J]. Peptides,2009,30 (4): 638-646.
[75]单晓枫,张洪波,郭伟生,等.乌鳢体表粘液抗菌肽CSM14的分离纯化及部分生物学活性研究[J].中国预防兽医学报,2010,32(3)：210-213.
[76]李伟,孙文秀,熊涛.黄鳝抗菌肽hepcidin基因cDNA的克隆及表达分析[J].湖南农业大学学报(自然科学版),2009,35(3)：304-307.
[77]Bao L B, Eric Peatman.The catfish liver-expressed antimicrobial peptide 2 (LEAP-2) gene is expressed in a wide range of tissues and developmentally regulated[J]. Molecular Immunology,2006,43 (4): 367-377.
[78]Salerno G, Parrinello N, Roch P. cDNA sequence and tissue expression of an antimicrobial peptide, dicentracin; a new component of the moronecidin family isolated from head kidney leukocytes of sea bass, Dicentrarchus labrax [J]. Comp Biochem Physiol B Biochem Mol Biol,2007,146(4):521-529.
[79]Chang C I, Zhang Y A. Two cathelicidin genes are present in both rainbow trout (Oncorhynchus mykiss) and atlantic salmon (Salm osalar) [J]. Antimicrob. Agents Chemother,2006.50(1):185-195.
[80]Ren H L, Wang K J, Zhou H L, et al. Cloning and organisation analysis of a hepcidin-like gene and cDNA from Japan sea bass,Lateolabrax japonicus[J]. Fish Shellfish Immunol,2006,21(3):221-227.
[81]苏建明,雷红宇,黄平,等.草鱼肠道抗菌肽的提取及体外抑菌效果初步研究[J].湖南农业大学学报(自然科学版),2009,35(2)：162-165.
[82]Paul Y, Kassebaum C, Lazarovici P, et al. Translocation of acylated pardaxin into cells[J]. FEBS Letters, 1998,440(122):131-134.
[83]单晓枫,吴同垒,王贵生,等.水产动物抗菌肽基因异源表达研究进展[J].中国兽药杂志,2011, 45(11):34-36.
[84]Srinivasulu B, Syvitski R, Seo J K, et al. Expression, purification and structural characterization of recombinant hepcidin, an antimicrobial peptide identified in Japanese flounder, Paralichthys olivaceus [J]. Protein Expr Purif,2008,61(1):36-44.
[85]张虹,陶妍.斑点叉尾(?)(Ictalurus punctatus)肝脏表达的抗菌肽-2(LEAP-2)在E.coli中的融合表达[J].上海交通大学学报(农业科学版),2010,28(1)：70-75.
[86]Hong I P, Lee S J, Choi S G. Recombinant expression of human cat helicidin (hCAP18/LL-37) in Pichia pastoris [J]. Biotechnol Lett,2007,29(1):73-78.
[87]蔡晶晶,杨明,蔡灵,等.黑鲷抗菌肽hepcidin在毕赤酵母中的表达及其抗菌活性[J].厦门大学学报(自然科学版),2009,48(5)：738-743.
[88]李伟,陈松林.大菱鲆抗菌肽基因酵母表达载体的构建及表达[J].长江大学学报(自然科学版),2008,5(4)：61-66.
[89]Sarmasika A, Chen T T. Bactericidal activity of cecropin B and cecropin P1 expressed in fish cells (CHSE-214):application in controlling fish bacterial pathogens[J].Aquac,2003(220):183-194.
[90]Rex A D, Dregory W W. Enhanced bacterial disease resistance of transgenic Channel catfish Ictalurus punctatus processing cecropin genes[J]. Marine Biotech,2002(4):338-343.
[91]Xiang J H. Disease occurrence and control strategies of mariculture organisms[J].Ocean Press,2001:1-4.
[92]Bogdan C, Rllinghoff M, Diefenbach A.Reactive oxygen and reactive nitrogen intermediates in innate and pecificimmunity[J].Curr. Opin. Immunol,2000,12(1):64-76.
[93]Yu B P.Cellular defenses against damage from reactive oxygen species[J].Physiol. Rev,1994,74(1): 139-162.
[94]Phalgun J, Dennis P P. Struetural, function and evolytion of the family of superoxide dismutase proteins from Halophilic arhaebacteria[J].J Bacteriol,1990,175(6):1572-1580.
[95]Barry L S, Dagmar R, Gregory A P.The 2.1 A resolution strueture of iron superoxide dismutase from Pseudomonas ovalis[J].Biochemistry,1990,265(29):8885-8893.
[96]Lin Y C, Baskaralingam V, Chen J C. Identification of the extracellular copper zinc superoxide dismutase (ecCuZnSOD) gene of the mud crab Scylla serrata and its expression following β-glucan and peptidoglycan injections[J]. Molecular Immunology,2008,45:1346-1355.
[97]Tian J X, Chen J, Jiang D, et al. Transcriptional regulation of extracellular copper zinc superoxide dismutase from white shrimp Litopenaeus vannamei following Vibrio alginolyticus and WSSV infection[J]. Fish & Shellfish Immunology,2011,30:234-240.
[98]Cerenius L, Lee B L. The proPO-system:pros and cons for its role in invertebrate immunity[J]. Trends Immunol,2008,29:63-71.
[99]Go'mez-Anduro G A, Barillas-Mury C V, Peregrino-Urarte A B, et al. The cytosolic manganese superoxide dismutase from the shrimp Litopenaeus vannamei:molecular cloning and expression[J]. Dev Comp Immunol,2006,30:893-900.
[100]Nirmala P, Akira M, Sloane S, et al. Generation and characterization of a novel kidney-specific manganese superoxide dismutase knockout mouse[J]. Free Radical Biology & Medicine,2011,51: 406-416.
[101]Murphy MP. How mitochondria produce reactive oxygen species[J]. Biochemical Journal,2009,417: 1-13.
[102]Kuwahara H, Horie T, Ishikawa S, et al. Oxidative stress in skeletal Muscle causes severe disturbance of exercise activity without muscle atrophy[J]. Free Radical Biology and Medicine,2010,48:1252-1262.
[103]郭建军,龚兴国.钝顶螺旋藻Fe-SOD基因的克隆及序列分析[J].浙江大学学报(理学版),2004,31(6)：674-678.
[104]Drivas GT, Shih A, Coutavas E, et al.Charaeterization of four novel ras-like genes expressed in a human teratoeareinoma cell line[J]. Mol.Cell.Biol,1990,10(4):1793-1798.
[105]Kaibuehi K, Kuroda S, Anlano M.Regulation of the cytoskeleton and cell adhesion by the Rho family GTPases in manunalian cells[J].Annu Rev Bioehem,1999,68:459-486.
[106]Nobes C D, Hall A. Rho GTPases control polarity, protrusion, and adhesion during cell movement[J]. J of Cell Biology,1999,144:1235-1244.
[107]Mulloy J C, Caneelas J A, Filippi M D, et al.Rho GTPases in hematopoiesis and hemopathies[J]. Blood, 2010,115(5):936-947.
[108]Anne J.Ridley.Rho proteins:linking signaling with membrane trafficking[J]. Traffie, 2001,2:303-310.
[109]Terry S, Nie M, Matter K, et al.Rho signaling and tight junction functions[J].Physiology (Bethesda), 2010,25(1):16-26.
[110]Chen BC, Kang J C, Lu Y T, et al.Racl regulates Peptidoglyean-induced nuclear factor-kappaB activation and cyclooxygenase-2 expression in RAW 264.7 macrophages by activating the phosphatidylinositol 3-kinase/Akt pathway[J].Mol Immunol,2009,46(6)::1179-1188.
[111]George A, Pushkaran S, Li L, et al.Altered phosphorylation of cytoskeleton proteins in sickle red blood cells:the role of protein kinase C, Rac GTPases, and reactive oxygen species[J]. Blood Cells Mol Dis, 2010,45(1):41-45.
[112]Monypenny J, Zicha D, Higashida C, et al.Cdc42 and Rac family GTPases regulate mode and speed but not direction of primary fibroblast migration during platelet-derived growth factor dependent chemotaxis[J].Mol Cell Biol,2009,29(10):2730-2747.
[113]Hiramoto-Yamaki N, Takeuehi S, Ueda S, et al. Ephexin4 and EphA2 mediate cell migration through aRhoG-dependent mechanism[J].J Cell Biol,2010,190(3):461-477.
[114]Tybulewiez V L, Henderson R B.Rho family GTPases and their regulators in lymphoeytes[J]. Nat Rev Immunol,2009,9(9):630-644.
[115]Yamazaki D, Kurisu S, TakenawaT. Involvement of Rac and Rho signaling incancer cell motility in 3D substrates Rac regulates cancer cell motility in 3D ECMs[J].Oncogene,2009,28:1570-1583.
[116]Kami Schlessinger, Alan Hall. Nicholas Tolwinski.Wnt signaling pathways meet Rho GTPases[J]. Genes&Dev,2009,23:265-277.
[117]Johnson E, Seachrist D D, DeLeon-Rodriguez C M, et al.HER2/ErbB2-induced breast cancer cell migration and invasion require p120 catenin activation of Racl and Cdc42[J]. J Biol Chem,2010, 285(38):29491-29501.
[118]薛妍,毕锋,刘娜.Rho GTPases对肿瘤血管生成相关分子的作用[J].中国生物化学与分子生物学报,2004,20(5)：664-669.
[119]Tsai N P, Wei L N. RhoA/ROCK1 signaling regulates stress granule formation and apoptosis[J]. Cell Signal,2010,22(4):668-675.
[120]Matos P, Jordan P. Increased Raclb expression sustains colorectal tumor cell survival[J]. Mol Cancer Res,2008,6(7):1178-1184.
[121]Sengupta A, Amett J, Dunn S, et al. Rac2 GTPase deficiency depletes BCR-ABL+leukemic stem cells and progenitors in vivo[J]. Blood,2010,116(1):81-84.
[122]O Brien PJ. Chemico-Biological Interactions[J]. Peroxidases,2000,129:113-139.
[123]Podrez E A, Poliakov E, Shen Z, et al. A novel family of atherogenic oxidized phospholipids promotes macrophage foam cell formation via the scavenger receptor CD36 and is enriched in atherosclerotic lesions [J]. J Biol Chem,2002,277:38517-38523.
[124]Klebanoff S J.Myeloperoxidase:a friend and foe[J]. J Leukoeyte Biology,2005,77(5):598-625.
[125]汤红英,郑玉建,吴顺华.髓过氧化物酶相关疾病的研究进展[J].新疆医科大学学报,2009,32(3)：255-258.
[126]Bielek E. Developmental stages and loealization of peroxidase activity in leukoeytes of three teleost species[J].Cel and Tissue Researeh,1981,220(1):163-180.
[127]PalicD, Andreasen C B, Menzel B W, et al. A rapid,direct assay to measure degranulation of primary granules in neutrophils from kidney of fathead minnow (Pimephales promelas Rafinesque,1820) [J]. Fish Shellfish Immuno,2005,19(3):217-227.
[128]Palic D, Andreasen C B, Ostojic J, et al. Zebrafish(Danio rerio)whole kidney assays to measure neutrophil extracellular trap release and degranulation of primary granules[J]Journal of Immunological Methods,2007,319(1-2):87-97.
[129]Yoshiaki M, Akihiro Y, Yasuyuki S, et al. The identification of genes specific to Prevotella intermedia and Prevotella nigrescens using genomic subtractive hybridization[J]. Anaerobe,2010,16:265-269.
[130]Tachibana-Ono M, Yoshida A, Kataoka S, et al. Identification of the genes associated with a virulent strain of Porphyromonas gingivalis using the subtractive hybridization technique[J]. Oral Microbiol Immunol,2008,23:84-87.
[131]Zhang X, Zhou YL, Anne K. Isolation and characterization of novel pituitary tumor related genes:A cDNA representational difference approach[J]. Molecular and Cellular Endocrinology,2010,326:40-47.
[132]Blaes N, Elbaz M, Heitz F, et al. Differential display fingerprints:new approach to characterize smooth muscle cells and human coronary atherectomy tissues[J]. Pathologie Biologie,2007,55:328-335.
[133]Pascal P, Sebastien B, Thomas C, et al. Pitfalls of reverse transcription quantitative polymerase chain reaction standardization:Volume-related inhibitors of reverse transcription[J]. Analytical Biochemistry, 2011,415:151-157.
[134]Toshiaki W, Takashi K, Yoko Y, et al. Prediction of liver metastasis after colorectal cancer using reverse transcription-polymerase chain reaction analysis of 10 genes[J]. European j ournal of cancer,2010,46: 2119-2126.
[135]单晓枫,孙佳敏,张洪波,等.抑制性消减杂交(SSH)技术及其在水生动物基因克隆中的应用[J].黑龙江畜牧兽医,2010,12(上)：30-32.
[136]Diatchenko L, Lau Y F, Campbell A P, et al.Suppression subtractive hybridization:a method for generating differentially regulated or tissue-specific cDNA probes and libraries[J]. Proc Natl Acad Sci USA,1996,93:6025-6030.
[137]Rebrikov D V, Britanova O V, Gurskaya N G, et al. Mirror orientation selection(MOS):a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization[J]. Nucleic Acids Research,2000,28(20):91-94.
[138]Wan J, Matthew B W, Li C, et al. Efficacy of SSH PCR in isolating differentially expressed genes[J].B M C Genomics,2002,3(12):1-7.
[139]刘晓慧,王秀,王义权.文昌鱼性腺差减cDNA文库的构建[J].动物学报,2008,54(3)：482-488.
[140]Wang K J, Ren H L, Xu D D, et al. Identification of the up-regulated expression genes in hemocytes of variously colored abalone(Haliotis diversicolor Reeve,1846) challenged with bacteria[J]. Developmental and Comparative Immunology,2008,32:1326-1347.
[141]Zhao Z Y, Yin Z X, Weng S P, et al. Profiling of differentially expressed genes in hepatopancreas of white spot syndrome virus-resistant shrimp(Litopenaeus vannamei) by suppression subtractive hybridization [J].Fish Shellfish Immunology,2007,22(5):520-534.
[142]Lin B, Chen S W, Cao Z, et al. Acute phase response in zebrafish upon Aeromonas salmonicida and Staphylococcus aurens infection:Striking similarities and obvious differences with mammals [J]. Mol Immunol,2007,44:295-301.
[143]Griffitt R J, Chandler G T, Greig T W, et al.Cathepsin B and glutathione peroxidase show differing transcriptional responses in the grass shrimp, Palaemonetes pugio following exposure to three xenobiotics[J]. Environ Sci Technol,2006,40(11):3640-3645.
[144]Li Z, Yu M, Zhang H, et al. Improved rapid ampliation of cDNA ends (RACE) for mapping both the 5' and 3' terminal sequences of paramyxovirus genomes[J].Journal of Virological Methods, 2005,130: 154-156.
[145]Fernando L P, Peter L. A guide for in-house design of template-switch-based 50 rapid amplication of cDNA ends systems[J]. Analytical Biochemistry,2010,397:227-232.
[146]Inoue Y, Saito T, Endo M, et al. Molecular cloning and preliminary expression analysis of banded dogfish(Triakis cyllia) CC chemokine cDNAs by use of suppression subtractive hybridization[J]. Immunogenetics,2005,56:722-734.
[147]Tsutsui S, Iwamoto T, Nakamura O, et al. LPS induces gene expression of interleukin-1b in conger eel (Conger myriaster) macrophages:First cytokine sequence within Anguilliformes[J].Fish Shellfish Immunology,2007,23:911-916.
[148]贾伟章,周秀霞,黄容,等.乌鳢(Channa argus)干扰素刺激基因Viperin和ISG15及其启动子的克隆与特征分析[J].自然科学进展,2007,17(11)：1841-1941.
[149]Jia W, Guo Q. Gene structures and promoter characteristics of interferon regulatory factor 1 (IRF-1), IRF-2 and IRF-7 from snakehead Channa argus[J]. Mol Immunol,2008,45(8):2419-2428.
[150]徐田军,陈松林.鱼类MHC及其基因研究进展[J].海洋水产研究,2008,29(4)：116-126.
[151]池磊,刘扬,冯琳,等.鱼类组蛋白衍生抗菌肽与其抗菌的关系[J].2009,30(6)：56-60.
[152]Huai L F, Jay I S, Amy E T. Decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility[J]. Fertility and Sterility,2001,76(6):1136-1139.
[153]Jeanne B, John P, Karthish M, et al. Comparing the functional properties of the Hsp70 chaperones, DnaK and BiP[J]. Biophysical Chemistry,2010,149:58-66.
[154]张永安,聂品,朱作言.鱼类免疫球蛋白基因结构及抗体多样性的遗传机制[J].遗传,2002,24(5)：575-580.
[155]肖凡书,聂品.鱼类免疫球蛋白重链基因与基因座的研究进展[J].水产学报,2010,34(10)：1617-1682.
[156]杜浩,危起伟,杨德国,等.MS- 222、丁香油、苯唑卡因对养殖美洲鲥幼鱼的麻醉效果[J].大连水产学院学报,2007,22(1)：20-26.
[157]梁政远,安丽娜,缪凌鸿,等.丁香油对鲤鱼和罗非鱼的麻醉作用[J].水产学杂志,2009,22(1)：42-46.
[158]张立颖,赵萌,王跃智.水生生物超氧化物岐化酶的研究进展[J].江西农业大学学报,2012,34(4)：800-804.
[159]Cho Y S, Choi B N, Kim K H, et al.Differential expression of Cu/Zn superoxide dismutase mRNA during exposures to heavy metals in rockbream(OPlenathus faseiatus)[J].Aquaeulture,2006,253:667-679.
[160]Zhang Z W, Li Z, Liang H W, et al.Molecular cloning and differential expression pattern of copper/zinc superoxide dismutase and manganese superoxide dismutase in hypophthalmichthys molitrix[J]. Fish Shellfish Immunology,2011,30:473-479.
[161]Lushehak V I, Bagnyukova T V. Effeets of different environmental oxygen levels on free radical processes in fish[J].Comp Biochem physiol Part,2006,144:283-289.
[162]Wennerberg K, Der CJ. Rho-family GTPases:it's not only Rac and Rho (and I like it) [J]. J Cell Sci 2004,117:1301-1312.
[163]Didsbury J, Weber RF, Bokoch GM, et al. rac, a novel rasrelated family of proteins that are botulinum toxin substrates[J]. J Biol Chem 1989,264:16378-16382.
[164]Haataja L, Groffen J, Heisterkamp N. Characterization of RAC3, a novel member of the Rho familyfJ]. J Biol Chem,1997,272:20384-20388.
[165]Williams D A, Tao W, Yang F, et al. Dominant negative mutation of the hematopoietic-specific Rho GTPase, Rac2, is associated with a human phagocyte immunodeficiency[J]. Blood,2000,96:1646-1654.
[166]Schmidt A, Hall A.Guanine nucleotide exchange factors for Rho GTPases:turing on the switch[J].Genes Dev,2002,16:1587-1609.
[167]Ambruso D R, Knall C, Abell A N, et al.Human neutrophil immunodeficiency syndrome is associated with an inhibitory Rce2 mutation[J]. Proc Natl Acad Sci USA,2000,97:4654-4659.
[168]Deng Q, Yoo S K, Cavnar PJ, et al. Dual roles for rac2 in neutrophil motility and active retention in zebrafish hematopoietic tissue[J].Dev Cell,2011,21:735-745.
[169]Tell R M, Kimura K, Palic D. Rac2 expression and its role in neutrophil functions of zebrafish (Danio rerio) [J]. Fish Shellfish Immunology,2012,33:1086-1094.
[170]Roberts A W, Kim C, Zhen L, et al. Deficiency of the hematopoietic cell-specific Rho family GTPase Rac2 is characterized by abnormalities in neutrophil function and host defense[J].Immunity,1999,10: 183-196.
[171]Abdel-Latif D, Steward M, Macdonald D L, et al. Rac2 is critical for neutrophil primary granule exocytosis[J]. Blood,2004,104:832-839.
[172]Jia A, Zhang X H.cDNA cloning, characterization, and expression analysis of the Racl gene from Scophthalmus maximus[J]. Com Biochem PhyB,2009,154(1):80-84.
[173]Jantschko W, Georg F P, Zederbauer M, et al. Direct conversion of ferrous myeloperoxidase to compound Ⅱ by hydrogen peroxide:an anaerobic stopped-flow study[J]. Biochemical and Biophysical Research Communications,2003,312(2):292-298.
[174]Bradley P P, Priebat D A, Christensen R D, et al.Measurement of cutaneous inflammation estimation of neutrophil content with an enzyme marker [J]. Journal of Investigative Dermatology,1982,78(3): 206-209.
[175]Lieschke G J, Oates A C, Crowhurst M O, et al.Morp hologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish [J]. Blood,2001,98(10):3087-3096.
[176]Bermett C M, Kanki J P, Rhodes J, et al.Myelopoiesis in the zebrafish, Danio rerio[J].Blood,2001, 98(3):643-651.
[177]胡宝庆,刘毅,文春根.翘嘴鳜髓过氧化物酶基因克隆及重链序列的原核表达[J].山东大学学报(理学版),2009,44(5)：10-19.
[178]Yeh H Y, Klesius P H.Sequence analysis,charaeterization and tissue distribution of channelcatfish (Ictalurus punctatus Rafinesque,1818) myeloperoxidase cDNA[J].Fish shellfish Immunology,2010, 28(3):504-509.
[179]Johnson K R, Nauseef W M, Care A, et al. Characterization of cDNA clones for human myeloperoxidase: predicted amino acid sequence and evidence for multiple mRNA species [J].Nucleic Acids Research, 1987,15(5):2013-2028.
[180]Morishita K, Kubota N, Asano S, et al. Molecular cloning and characterization of cDNA for human myeloperoxidase[J]. The Journal of Biological Chemistry,1987,262(8):3844-3851.
[181]Zeng J, Fenna R E. X-ray crystal structure of canine myeloperoxidase at 3 A resolution[J]. Journal of Molecular Biology,1992,226(1):185-207.
[182]Furtmuller P G, Zederbaue M, Jantschko W, et al. Active site structure and catalytic mechanisms of human peroxidases[J]. Archives of Biochemistry and Biophysics,2006,445(2):199-213.
[183]朱元晓,赖春宁,王建安,等.人髓过氧化物酶基因cDNA克隆及急性白血病髓过氧化物酶基因的表达[J].生物工程学报,1993,9(1)：21-24.
[2]王广军.乌鳢的生物学特性及繁殖技术[J].淡水渔业,2000,30(6)：10-11.
[3]聂国兴,傅艳茹,张浩,等.乌鳢肌肉营养成分分析[J].淡水渔业,2002,32(2)：46-47.
[4]吴同垒,单晓枫,王桂芹,等.乌鳢主要病害研究进展[J].黑龙江畜牧兽医,2011,9：29-31.
[5]潘立新,郑善坚.乌鳢暴发性出血病的诊断和治疗[J].淡水渔业,2004，34(4)：49-50.
[6]曹铮,沈锦玉,谢荣林,等.乌鳢结节病病原-诺卡氏菌的分离与鉴定[J].水生生物学报,2009,33(2)：338-340.
[7]黄江峰,饶毅,金友萍,等.乌鳢溃疡综合症流行病规律调查[J].江西水产科技,2008,4：37-39.
[8]张海宾.乌鳢出血性败血症病原研究[J].水利渔业,2005,25(2)：81-82.
[9]舒新华,金燮理,肖克宇,等.乌鳢腹水病病原的分离和鉴定[J].湖南农业大学学报,1998,24(4)：286-290.
[10]曾慷,舒新华,金燮理.乌鳢腐皮病致病细菌的研究[J].湖南农业大学学报,1999,25(5)：396-399.
[11]舒新华,曾慷,金燮理,等.乌鳢腐皮病病理形态学研究[J].淡水渔业,1999,29(8)：8-10.
[12]袁思平,王国良,金珊.养殖鱼类致病诺卡氏菌研究进展[J].微生物通报,2006,33(2)：137-141.
[13]王家沛.乌鳢诺卡氏菌病的诊治[J].现代农业科技,2010(13)：387-390.
[14]吕伟清,孙伟杰,谢荣林.乌鳢诺卡氏菌的防治[J].科学养鱼,2008(1)：57.
[15]王国良,徐益军,金珊,等.养殖乌鳢诺卡氏菌病及其病原研究[J].水生生物学报,2009,33(2)：277-283.
[16]石存斌,潘厚军,常藕琴,等.养殖乌鳢结节病的病原分析[J].安徽农业科学,2009,37(16)：7384-7386.
[17]胡银亨.乌鳢车轮虫研究[J].泸州职业技术学院学报,2007,4：34-37.
[18]Mauel M J, Miller D L. Piscirickettsiosis and piscirickettsiosis-like infections in fish:a review[J].Vet Microbiol,2002,87:279-289.
[19]贾伟章,孙晓凤,郭琼林.养殖乌鳢类立克次体分离纯化的初步研究[J].水生生物学学报,2004,28(4)：448-449.
[20]Guo Q L, Jia W Z, Han X P, et al. Rickettsia-like organism infection in a freshwater cultured fish Ophiocephalus argus C. in China [J]. Prog Nat Sci.2004,14:417-422.
[21]单晓枫,郭伟生,张洪波,等.鱼类体液中的几种抗菌因子研究进展[J].河南农业科学,2010,5：138-142.
[22]Claire M H, Lambris J D. The complement system in teleosts [J].Fish&Shellfish Immunology,2002, 12:399-420.
[23]Kuroda N, Naruse K, Shima A, et al. Molecular cloning and linkage analysis of comp lement C3 and C4 genes of the Japanese medakafish [J]. Immunogenetics.2000,51:117-128.
[24]NakaoM, Mutsuro J, Obo R, et al. Molecular cloning and protein analysis of divergent forms of the complement component C3 from a bony fish, the common carp (Cyprinus carpio):presence of variants lacking the catalytic histidine[J]. Eur J Immunol,2000,30:858-866.
[25]Smith S H, Jensen J A. The second component (C2n) of the nurse shark complement system:purification, physico-chemical characterization and functional comparison with guinea pig C4 [J]. Dev Comp Immunol,1986,10:191-206.
[26]Terado T, Okamura K, Ohta Y, et al. Molecular cloning of C4 gene and identification of the classIII complement region in the shark MHC [J]. J Immunol,2003,171:2461-2466.
[27]Shin D H, Webb B, NakaoM, et al. Molecular cloning, structural analysis and expression of complement component B f/C2 genes in the nurse shark, Ginglym ostom a cirratum[J]. Dev Comp Immunol,2007,, 31:1168-1182.
[28]Wang T, Secombes C J. Complete sequencing and expression of three complement components, Clr, C4 and Cl inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss [J]. Immunogenetics,2003,55:615-628.
[29]Ellingsen T, Strand C, Monsen E, et al. The ontogeny of complement component C3 in the spotted wolfish (Anarhichasminor olafsen) [J]. Fish Shellfish Immunol,2005,18:351-358.
[30]Lange S, Slavko H B, Alister W D, et al. Complement component C3 transcription in Atlantic halibut (Hippoglossus hippoglossus L.) larvae [J]. Fish Shellfish Immunol,2006,20:285-294.
[31]Marie Lovoll, Terje Kilvik, HaniBoshra, et al. Maternal transfer of complement components C3-1, C3-3, C3-4, C4, C5, C7, Bf and Df to off spring in rainbow trout (Oncorhynchus mykiss) [J].Immunogenetics, 2006,58:168-179.
[32]Fujita T. Evolution of lecfin-complement pathway and its role in innate immunity [J]. Nature,2002,2: 347-353.
[33]Boshra H, Li J, Sunyer J O. Recent advances on the complement system of teleost fish [J]. Fish Shellfish Immunol,2006,20:239-262.
[34]Boshra H, Gelman A E, Sunyer J O. Structural and functional characterization of complement C4 and Cls-like molecules in teleost fish:insights into the evolution of classical and alternative pathways [J]. J Immunol,2004,173:349-359.
[35]Endo Y, Takahashi M, Fujita T. Lectin complement system and pattern recognition [J]. Immunolobiology,2006,211:283-293.
[36]Boshra H, Peters R, Li J, et al. Production of recombinant C5a from rainbow trout (Oncorhynchus mykiss):role in leucocyte chemotaxis and respiratory burst [J]. Fish Shellfish Immunol,2004,17: 293-303.
[37]Barnes A C, Ellis A E. Role of capsule in serotypic differences and complement fixation by lactococcus garvieae[J]. Fish Shellfish Immunol,2004,16:207-214.
[38]Fernandez-Trujillo M A, Bejar J, Gallardo J B, et al. Molecular cloning and characterization of C-type lysozyme from Senegalese sole (Solea senegalensis) [J]. Aquaculture,2007,272 (Supp 11):255.
[39]Fast M D, Sims D E, Burka J F, et al. Skin morphology and humoral non-specific defence parameters of mucus and plasma in rainbow trout, coho and Atlantic salmon[J]. Comp Biochem Physiol A Mol Integr Physiol,2002,132(3):645-657.
[40]刘红柏,魏巍.史氏鲟及小体鲟不同组织中溶菌酶水平的比较[J].水产学杂志,2007,20(1)：57-60.
[41]Subbotkina TA, Subbotkin M F. Lysozyme content in organs and blood serum in various species in the Volga River [J]. J. Evolutionary Biochemistry and Physiology,2003,39 (5):537-546.
[42]Hikima J, Minagawa S, Hirono I, et al. Molecular cloning, expression and evolution of the Japnanese flounder goose-type lysozyme gene, and the lytic activity of its recombinant protein[J]. Biochimica et Biophysica Acta,2001,1520:35-44.
[43]Mitra A, Foster-Frey J, Rexroad C E, et al. Molecular characterization of lysozyme type Ⅱ gene in rainbow trout (Oncorhynchus mykiss):evidence of gene duplication[J].Anim Biotechnol,2003,14 (1):7-12.
[44]Yin Z X, He J G, Deng W X, et al. Molecular cloning, expression of orange-spotted grouper goose-type lysozyme cDNA, and lytic activity of its recombinant protein[J]. Dis Aquat Organ,2003,55 (2):117-123.
[45]纪伟,张培军.转“全鱼”溶菌酶基因大菱鲆的研究[J].海洋科学,2004,28(5)：8-14.
[46]Cho Y S, Lee S Y, Kim K Y, et al. Gene structure and expression of metallot hionein during metal exposures in Hemibarbus mylodon [J]. Ecotoxicol Environ Safety,2008,71(1):125-137.
[47]Schmidt C, Beyersmann D. Transient peaks in zinc and metalliothionein levels during differentiation of 3T3L1 cells [J].Arch Biochem Biophys,1999,364:91-98.
[48]Petrosillo G, Di Venosa N, Ruggiero F M, et al. Mitochondrial dysfunction associated wit h cardiac ischemia/reperfusion can be attenuated by oxygen tension control. Role of oxygenfree radicals and cardiolipin[J]. Biochim Biophy Acta Bioenerget,2005,1710(2-3):78-86.
[49]Haq F, Mahoney M, Koropatnick J. Signaling events for metallothionein induction [J]. Mutat Res,2003, 533:211-226.
[50]Bonhan k M, Zafarullahl, Gedamu. The rainbow trout metallothioneins molecular cloning and characterization of two distinct cDNA sequences [J]. DNA,1987, (6):519-528.
[51]赵晓祥,张淑梅,李晶,等.虹鳟鱼金属硫蛋白启动子的体外扩增、克隆及其序列分析[J].生物技术,2000,10(1)：1-3.
[52]冯仁勇,周小秋.鱼类金属硫蛋白基因的表达与调控[J].水产科学,2005,24(9)：52-54.
[53]龙华,曾勇.鱼血清转铁蛋白对嗜水气单胞菌的抗性研究[J].湖北农学院学报,2004,24(2)：119-123.
[54]FerixenetN, Remacha A, Berlanga E, et al. Serum soluble transferrin receptor concentrations are increased in central obesity. Results from a screening programme for hereditary hemochromatosis in men with hyperferritinemia[J]. Clinica Chimica Acta,2009,400:111-116.
[55]Park I, Schaeffer E, Sidoli A, et al. Organization of the human transferrin gene:Direct evidence that it originated by gene duplication[J]. Proc Natl Acad Sci U S A,1985,82:3149.
[56]Bartfeld N S, Law J H. Isolation and molecular cloning of transferring from the tobacco hornworm, Manduca Sexta[J].J Biol Chem,1990,265:21684-21691.
[57]Hirono I, Uchiyama T, Aoki T. Cloning, nucleotide sequence analysis, and characterization of cDNA for medaka (Oryzias latipes) transferring [J]. J Mar Biotechnol,1995,2:193-198.
[58]Lee J Y, Tange N, Yamashita H, et al. Cloning and characterization of transferrin cDNA from coho salmon (Oncorhynchus kisutch) [J]. Fish Pathology,1995,30(4):271-277.
[59]龙华.鲫血清转铁蛋白cDNA克隆及其序列分析[J].中山大学学报(自然科学版),2003,42(增刊)：231-234.
[60]李明云,陈炯,张祖兴,等.大黄鱼(Pseudosciaena crocea)转铁蛋白(Tf)基因克隆及序列分析[J].海洋与湖沼,2007,38(6)：563-567.
[61]林亚秋,郑玉才,吉红,等.草鱼转铁蛋白受体cDNA序列克隆及其组织表达差异[J].湖北农业科学,2009,48(6)：1289-1292.
[62]Fock W L, Chen C L, Lam TJ, et al. Roles of an endogenous serum lectin in immune protection of blue gourami, Trichogaster trichopteurs (Pallus) against Aeromonas hydrophila [J]. Fish Shellfish Immunol, 2001,11:245-252.
[63]Ottinger C A, Johnson S C, Ewart KV, et al. Enhancement of anti-Aeromonas salmonicida activity in Atlantic salmon (Salmo salar) macrophages by a mannose binding lectin [J].Comp Biochem Physiol C, 1999,123:53-59.
[64]Stratton L, WuS, Richards R C, et al. Oligomerisation and carbohydrate binding in an Atlantic salmon serum C-type lectin consistent with non-self recognition[J]. Fish Shellfish Immunol,2004,17:315-323.
[65]Okamoto M, Tsutsui S, Tasumi S, et al. Tandem repeat L-rhamnosebinding lectin from the skin mucus of ponyfish,Leiognathus nuchalis[J]. Biochem Bioph Res Co,2005,333:463-469.
[66]Jungw K, Park P J, Kim S K. Purification and characterization of a new lectin from the hard roe of skipjack tuna, Katsuwonus pelam is [J]. Int J Biochem Cell B,2003,35:255-265.
[67]吴超,陆承平.二龄草鱼脾脏、肝脏组织高表达甘露糖结合凝集素mRNA[J]动物学报,2004,50(1)：137-140
[68]Vitved L, Holmskov U, Koch C, et al. The homologue of mannose-binding lectin in the carp family Cyprinidae is expressed at high level in spleen, and deduced primary structure predicts affinity for galactose[J]. Immune-genetics,2000,51:955-964.
[69]吴超,陆承平.草鱼等8种淡水鱼血清凝集素作用的比较[J].南京农业大学学报,2004,27(2)：79-82.
[70]张黎,刘莉,袁媛,等.中华倒刺(?)血清凝集素的研究[J].南方水产,2006,2(6)：39-44.
[71]盘赛昆,顾小红,汤坚,等.鳙鳃凝集素的稳定性及细胞凝集作用[J].安徽农业科学,2009,37(26)：12589-12591.
[72]Huang P, Chen J, Kuo C.Three different hepcidins from tilapia,oreochromis mossambicus:Analysis of their expressions and biological functions[J]. Molecular Immunolgy,2007,44(8):1922-1934.
[73]Cuesta A, Meseguer J, Esteban M. The antimicrobial peptide hepcidin exerts an important role in the innate immunity against bacteria in the bony fish gilthead seabream [J]. Mol Immunol,2008, 45: 2333-2342.
[74]Wang KJ, Cai J J, Cai L, et al. Cloning and expression of a hepcidin gene from a marine fish (Pseudosciaena crocea) and the antimicrobial activity of it s synthetic peptide[J]. Peptides,2009,30 (4): 638-646.
[75]单晓枫,张洪波,郭伟生,等.乌鳢体表粘液抗菌肽CSM14的分离纯化及部分生物学活性研究[J].中国预防兽医学报,2010,32(3)：210-213.
[76]李伟,孙文秀,熊涛.黄鳝抗菌肽hepcidin基因cDNA的克隆及表达分析[J].湖南农业大学学报(自然科学版),2009,35(3)：304-307.
[77]Bao L B, Eric Peatman.The catfish liver-expressed antimicrobial peptide 2 (LEAP-2) gene is expressed in a wide range of tissues and developmentally regulated[J]. Molecular Immunology,2006,43 (4): 367-377.
[78]Salerno G, Parrinello N, Roch P. cDNA sequence and tissue expression of an antimicrobial peptide, dicentracin; a new component of the moronecidin family isolated from head kidney leukocytes of sea bass, Dicentrarchus labrax [J]. Comp Biochem Physiol B Biochem Mol Biol,2007,146(4):521-529.
[79]Chang C I, Zhang Y A. Two cathelicidin genes are present in both rainbow trout (Oncorhynchus mykiss) and atlantic salmon (Salm osalar) [J]. Antimicrob. Agents Chemother,2006.50(1):185-195.
[80]Ren H L, Wang K J, Zhou H L, et al. Cloning and organisation analysis of a hepcidin-like gene and cDNA from Japan sea bass,Lateolabrax japonicus[J]. Fish Shellfish Immunol,2006,21(3):221-227.
[81]苏建明,雷红宇,黄平,等.草鱼肠道抗菌肽的提取及体外抑菌效果初步研究[J].湖南农业大学学报(自然科学版),2009,35(2)：162-165.
[82]Paul Y, Kassebaum C, Lazarovici P, et al. Translocation of acylated pardaxin into cells[J]. FEBS Letters, 1998,440(122):131-134.
[83]单晓枫,吴同垒,王贵生,等.水产动物抗菌肽基因异源表达研究进展[J].中国兽药杂志,2011, 45(11):34-36.
[84]Srinivasulu B, Syvitski R, Seo J K, et al. Expression, purification and structural characterization of recombinant hepcidin, an antimicrobial peptide identified in Japanese flounder, Paralichthys olivaceus [J]. Protein Expr Purif,2008,61(1):36-44.
[85]张虹,陶妍.斑点叉尾(?)(Ictalurus punctatus)肝脏表达的抗菌肽-2(LEAP-2)在E.coli中的融合表达[J].上海交通大学学报(农业科学版),2010,28(1)：70-75.
[86]Hong I P, Lee S J, Choi S G. Recombinant expression of human cat helicidin (hCAP18/LL-37) in Pichia pastoris [J]. Biotechnol Lett,2007,29(1):73-78.
[87]蔡晶晶,杨明,蔡灵,等.黑鲷抗菌肽hepcidin在毕赤酵母中的表达及其抗菌活性[J].厦门大学学报(自然科学版),2009,48(5)：738-743.
[88]李伟,陈松林.大菱鲆抗菌肽基因酵母表达载体的构建及表达[J].长江大学学报(自然科学版),2008,5(4)：61-66.
[89]Sarmasika A, Chen T T. Bactericidal activity of cecropin B and cecropin P1 expressed in fish cells (CHSE-214):application in controlling fish bacterial pathogens[J].Aquac,2003(220):183-194.
[90]Rex A D, Dregory W W. Enhanced bacterial disease resistance of transgenic Channel catfish Ictalurus punctatus processing cecropin genes[J]. Marine Biotech,2002(4):338-343.
[91]Xiang J H. Disease occurrence and control strategies of mariculture organisms[J].Ocean Press,2001:1-4.
[92]Bogdan C, Rllinghoff M, Diefenbach A.Reactive oxygen and reactive nitrogen intermediates in innate and pecificimmunity[J].Curr. Opin. Immunol,2000,12(1):64-76.
[93]Yu B P.Cellular defenses against damage from reactive oxygen species[J].Physiol. Rev,1994,74(1): 139-162.
[94]Phalgun J, Dennis P P. Struetural, function and evolytion of the family of superoxide dismutase proteins from Halophilic arhaebacteria[J].J Bacteriol,1990,175(6):1572-1580.
[95]Barry L S, Dagmar R, Gregory A P.The 2.1 A resolution strueture of iron superoxide dismutase from Pseudomonas ovalis[J].Biochemistry,1990,265(29):8885-8893.
[96]Lin Y C, Baskaralingam V, Chen J C. Identification of the extracellular copper zinc superoxide dismutase (ecCuZnSOD) gene of the mud crab Scylla serrata and its expression following β-glucan and peptidoglycan injections[J]. Molecular Immunology,2008,45:1346-1355.
[97]Tian J X, Chen J, Jiang D, et al. Transcriptional regulation of extracellular copper zinc superoxide dismutase from white shrimp Litopenaeus vannamei following Vibrio alginolyticus and WSSV infection[J]. Fish & Shellfish Immunology,2011,30:234-240.
[98]Cerenius L, Lee B L. The proPO-system:pros and cons for its role in invertebrate immunity[J]. Trends Immunol,2008,29:63-71.
[99]Go'mez-Anduro G A, Barillas-Mury C V, Peregrino-Urarte A B, et al. The cytosolic manganese superoxide dismutase from the shrimp Litopenaeus vannamei:molecular cloning and expression[J]. Dev Comp Immunol,2006,30:893-900.
[100]Nirmala P, Akira M, Sloane S, et al. Generation and characterization of a novel kidney-specific manganese superoxide dismutase knockout mouse[J]. Free Radical Biology & Medicine,2011,51: 406-416.
[101]Murphy MP. How mitochondria produce reactive oxygen species[J]. Biochemical Journal,2009,417: 1-13.
[102]Kuwahara H, Horie T, Ishikawa S, et al. Oxidative stress in skeletal Muscle causes severe disturbance of exercise activity without muscle atrophy[J]. Free Radical Biology and Medicine,2010,48:1252-1262.
[103]郭建军,龚兴国.钝顶螺旋藻Fe-SOD基因的克隆及序列分析[J].浙江大学学报(理学版),2004,31(6)：674-678.
[104]Drivas GT, Shih A, Coutavas E, et al.Charaeterization of four novel ras-like genes expressed in a human teratoeareinoma cell line[J]. Mol.Cell.Biol,1990,10(4):1793-1798.
[105]Kaibuehi K, Kuroda S, Anlano M.Regulation of the cytoskeleton and cell adhesion by the Rho family GTPases in manunalian cells[J].Annu Rev Bioehem,1999,68:459-486.
[106]Nobes C D, Hall A. Rho GTPases control polarity, protrusion, and adhesion during cell movement[J]. J of Cell Biology,1999,144:1235-1244.
[107]Mulloy J C, Caneelas J A, Filippi M D, et al.Rho GTPases in hematopoiesis and hemopathies[J]. Blood, 2010,115(5):936-947.
[108]Anne J.Ridley.Rho proteins:linking signaling with membrane trafficking[J]. Traffie, 2001,2:303-310.
[109]Terry S, Nie M, Matter K, et al.Rho signaling and tight junction functions[J].Physiology (Bethesda), 2010,25(1):16-26.
[110]Chen BC, Kang J C, Lu Y T, et al.Racl regulates Peptidoglyean-induced nuclear factor-kappaB activation and cyclooxygenase-2 expression in RAW 264.7 macrophages by activating the phosphatidylinositol 3-kinase/Akt pathway[J].Mol Immunol,2009,46(6)::1179-1188.
[111]George A, Pushkaran S, Li L, et al.Altered phosphorylation of cytoskeleton proteins in sickle red blood cells:the role of protein kinase C, Rac GTPases, and reactive oxygen species[J]. Blood Cells Mol Dis, 2010,45(1):41-45.
[112]Monypenny J, Zicha D, Higashida C, et al.Cdc42 and Rac family GTPases regulate mode and speed but not direction of primary fibroblast migration during platelet-derived growth factor dependent chemotaxis[J].Mol Cell Biol,2009,29(10):2730-2747.
[113]Hiramoto-Yamaki N, Takeuehi S, Ueda S, et al. Ephexin4 and EphA2 mediate cell migration through aRhoG-dependent mechanism[J].J Cell Biol,2010,190(3):461-477.
[114]Tybulewiez V L, Henderson R B.Rho family GTPases and their regulators in lymphoeytes[J]. Nat Rev Immunol,2009,9(9):630-644.
[115]Yamazaki D, Kurisu S, TakenawaT. Involvement of Rac and Rho signaling incancer cell motility in 3D substrates Rac regulates cancer cell motility in 3D ECMs[J].Oncogene,2009,28:1570-1583.
[116]Kami Schlessinger, Alan Hall. Nicholas Tolwinski.Wnt signaling pathways meet Rho GTPases[J]. Genes&Dev,2009,23:265-277.
[117]Johnson E, Seachrist D D, DeLeon-Rodriguez C M, et al.HER2/ErbB2-induced breast cancer cell migration and invasion require p120 catenin activation of Racl and Cdc42[J]. J Biol Chem,2010, 285(38):29491-29501.
[118]薛妍,毕锋,刘娜.Rho GTPases对肿瘤血管生成相关分子的作用[J].中国生物化学与分子生物学报,2004,20(5)：664-669.
[119]Tsai N P, Wei L N. RhoA/ROCK1 signaling regulates stress granule formation and apoptosis[J]. Cell Signal,2010,22(4):668-675.
[120]Matos P, Jordan P. Increased Raclb expression sustains colorectal tumor cell survival[J]. Mol Cancer Res,2008,6(7):1178-1184.
[121]Sengupta A, Amett J, Dunn S, et al. Rac2 GTPase deficiency depletes BCR-ABL+leukemic stem cells and progenitors in vivo[J]. Blood,2010,116(1):81-84.
[122]O Brien PJ. Chemico-Biological Interactions[J]. Peroxidases,2000,129:113-139.
[123]Podrez E A, Poliakov E, Shen Z, et al. A novel family of atherogenic oxidized phospholipids promotes macrophage foam cell formation via the scavenger receptor CD36 and is enriched in atherosclerotic lesions [J]. J Biol Chem,2002,277:38517-38523.
[124]Klebanoff S J.Myeloperoxidase:a friend and foe[J]. J Leukoeyte Biology,2005,77(5):598-625.
[125]汤红英,郑玉建,吴顺华.髓过氧化物酶相关疾病的研究进展[J].新疆医科大学学报,2009,32(3)：255-258.
[126]Bielek E. Developmental stages and loealization of peroxidase activity in leukoeytes of three teleost species[J].Cel and Tissue Researeh,1981,220(1):163-180.
[127]PalicD, Andreasen C B, Menzel B W, et al. A rapid,direct assay to measure degranulation of primary granules in neutrophils from kidney of fathead minnow (Pimephales promelas Rafinesque,1820) [J]. Fish Shellfish Immuno,2005,19(3):217-227.
[128]Palic D, Andreasen C B, Ostojic J, et al. Zebrafish(Danio rerio)whole kidney assays to measure neutrophil extracellular trap release and degranulation of primary granules[J]Journal of Immunological Methods,2007,319(1-2):87-97.
[129]Yoshiaki M, Akihiro Y, Yasuyuki S, et al. The identification of genes specific to Prevotella intermedia and Prevotella nigrescens using genomic subtractive hybridization[J]. Anaerobe,2010,16:265-269.
[130]Tachibana-Ono M, Yoshida A, Kataoka S, et al. Identification of the genes associated with a virulent strain of Porphyromonas gingivalis using the subtractive hybridization technique[J]. Oral Microbiol Immunol,2008,23:84-87.
[131]Zhang X, Zhou YL, Anne K. Isolation and characterization of novel pituitary tumor related genes:A cDNA representational difference approach[J]. Molecular and Cellular Endocrinology,2010,326:40-47.
[132]Blaes N, Elbaz M, Heitz F, et al. Differential display fingerprints:new approach to characterize smooth muscle cells and human coronary atherectomy tissues[J]. Pathologie Biologie,2007,55:328-335.
[133]Pascal P, Sebastien B, Thomas C, et al. Pitfalls of reverse transcription quantitative polymerase chain reaction standardization:Volume-related inhibitors of reverse transcription[J]. Analytical Biochemistry, 2011,415:151-157.
[134]Toshiaki W, Takashi K, Yoko Y, et al. Prediction of liver metastasis after colorectal cancer using reverse transcription-polymerase chain reaction analysis of 10 genes[J]. European j ournal of cancer,2010,46: 2119-2126.
[135]单晓枫,孙佳敏,张洪波,等.抑制性消减杂交(SSH)技术及其在水生动物基因克隆中的应用[J].黑龙江畜牧兽医,2010,12(上)：30-32.
[136]Diatchenko L, Lau Y F, Campbell A P, et al.Suppression subtractive hybridization:a method for generating differentially regulated or tissue-specific cDNA probes and libraries[J]. Proc Natl Acad Sci USA,1996,93:6025-6030.
[137]Rebrikov D V, Britanova O V, Gurskaya N G, et al. Mirror orientation selection(MOS):a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization[J]. Nucleic Acids Research,2000,28(20):91-94.
[138]Wan J, Matthew B W, Li C, et al. Efficacy of SSH PCR in isolating differentially expressed genes[J].B M C Genomics,2002,3(12):1-7.
[139]刘晓慧,王秀,王义权.文昌鱼性腺差减cDNA文库的构建[J].动物学报,2008,54(3)：482-488.
[140]Wang K J, Ren H L, Xu D D, et al. Identification of the up-regulated expression genes in hemocytes of variously colored abalone(Haliotis diversicolor Reeve,1846) challenged with bacteria[J]. Developmental and Comparative Immunology,2008,32:1326-1347.
[141]Zhao Z Y, Yin Z X, Weng S P, et al. Profiling of differentially expressed genes in hepatopancreas of white spot syndrome virus-resistant shrimp(Litopenaeus vannamei) by suppression subtractive hybridization [J].Fish Shellfish Immunology,2007,22(5):520-534.
[142]Lin B, Chen S W, Cao Z, et al. Acute phase response in zebrafish upon Aeromonas salmonicida and Staphylococcus aurens infection:Striking similarities and obvious differences with mammals [J]. Mol Immunol,2007,44:295-301.
[143]Griffitt R J, Chandler G T, Greig T W, et al.Cathepsin B and glutathione peroxidase show differing transcriptional responses in the grass shrimp, Palaemonetes pugio following exposure to three xenobiotics[J]. Environ Sci Technol,2006,40(11):3640-3645.
[144]Li Z, Yu M, Zhang H, et al. Improved rapid ampliation of cDNA ends (RACE) for mapping both the 5' and 3' terminal sequences of paramyxovirus genomes[J].Journal of Virological Methods, 2005,130: 154-156.
[145]Fernando L P, Peter L. A guide for in-house design of template-switch-based 50 rapid amplication of cDNA ends systems[J]. Analytical Biochemistry,2010,397:227-232.
[146]Inoue Y, Saito T, Endo M, et al. Molecular cloning and preliminary expression analysis of banded dogfish(Triakis cyllia) CC chemokine cDNAs by use of suppression subtractive hybridization[J]. Immunogenetics,2005,56:722-734.
[147]Tsutsui S, Iwamoto T, Nakamura O, et al. LPS induces gene expression of interleukin-1b in conger eel (Conger myriaster) macrophages:First cytokine sequence within Anguilliformes[J].Fish Shellfish Immunology,2007,23:911-916.
[148]贾伟章,周秀霞,黄容,等.乌鳢(Channa argus)干扰素刺激基因Viperin和ISG15及其启动子的克隆与特征分析[J].自然科学进展,2007,17(11)：1841-1941.
[149]Jia W, Guo Q. Gene structures and promoter characteristics of interferon regulatory factor 1 (IRF-1), IRF-2 and IRF-7 from snakehead Channa argus[J]. Mol Immunol,2008,45(8):2419-2428.
[150]徐田军,陈松林.鱼类MHC及其基因研究进展[J].海洋水产研究,2008,29(4)：116-126.
[151]池磊,刘扬,冯琳,等.鱼类组蛋白衍生抗菌肽与其抗菌的关系[J].2009,30(6)：56-60.
[152]Huai L F, Jay I S, Amy E T. Decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility[J]. Fertility and Sterility,2001,76(6):1136-1139.
[153]Jeanne B, John P, Karthish M, et al. Comparing the functional properties of the Hsp70 chaperones, DnaK and BiP[J]. Biophysical Chemistry,2010,149:58-66.
[154]张永安,聂品,朱作言.鱼类免疫球蛋白基因结构及抗体多样性的遗传机制[J].遗传,2002,24(5)：575-580.
[155]肖凡书,聂品.鱼类免疫球蛋白重链基因与基因座的研究进展[J].水产学报,2010,34(10)：1617-1682.
[156]杜浩,危起伟,杨德国,等.MS- 222、丁香油、苯唑卡因对养殖美洲鲥幼鱼的麻醉效果[J].大连水产学院学报,2007,22(1)：20-26.
[157]梁政远,安丽娜,缪凌鸿,等.丁香油对鲤鱼和罗非鱼的麻醉作用[J].水产学杂志,2009,22(1)：42-46.
[158]张立颖,赵萌,王跃智.水生生物超氧化物岐化酶的研究进展[J].江西农业大学学报,2012,34(4)：800-804.
[159]Cho Y S, Choi B N, Kim K H, et al.Differential expression of Cu/Zn superoxide dismutase mRNA during exposures to heavy metals in rockbream(OPlenathus faseiatus)[J].Aquaeulture,2006,253:667-679.
[160]Zhang Z W, Li Z, Liang H W, et al.Molecular cloning and differential expression pattern of copper/zinc superoxide dismutase and manganese superoxide dismutase in hypophthalmichthys molitrix[J]. Fish Shellfish Immunology,2011,30:473-479.
[161]Lushehak V I, Bagnyukova T V. Effeets of different environmental oxygen levels on free radical processes in fish[J].Comp Biochem physiol Part,2006,144:283-289.
[162]Wennerberg K, Der CJ. Rho-family GTPases:it's not only Rac and Rho (and I like it) [J]. J Cell Sci 2004,117:1301-1312.
[163]Didsbury J, Weber RF, Bokoch GM, et al. rac, a novel rasrelated family of proteins that are botulinum toxin substrates[J]. J Biol Chem 1989,264:16378-16382.
[164]Haataja L, Groffen J, Heisterkamp N. Characterization of RAC3, a novel member of the Rho familyfJ]. J Biol Chem,1997,272:20384-20388.
[165]Williams D A, Tao W, Yang F, et al. Dominant negative mutation of the hematopoietic-specific Rho GTPase, Rac2, is associated with a human phagocyte immunodeficiency[J]. Blood,2000,96:1646-1654.
[166]Schmidt A, Hall A.Guanine nucleotide exchange factors for Rho GTPases:turing on the switch[J].Genes Dev,2002,16:1587-1609.
[167]Ambruso D R, Knall C, Abell A N, et al.Human neutrophil immunodeficiency syndrome is associated with an inhibitory Rce2 mutation[J]. Proc Natl Acad Sci USA,2000,97:4654-4659.
[168]Deng Q, Yoo S K, Cavnar PJ, et al. Dual roles for rac2 in neutrophil motility and active retention in zebrafish hematopoietic tissue[J].Dev Cell,2011,21:735-745.
[169]Tell R M, Kimura K, Palic D. Rac2 expression and its role in neutrophil functions of zebrafish (Danio rerio) [J]. Fish Shellfish Immunology,2012,33:1086-1094.
[170]Roberts A W, Kim C, Zhen L, et al. Deficiency of the hematopoietic cell-specific Rho family GTPase Rac2 is characterized by abnormalities in neutrophil function and host defense[J].Immunity,1999,10: 183-196.
[171]Abdel-Latif D, Steward M, Macdonald D L, et al. Rac2 is critical for neutrophil primary granule exocytosis[J]. Blood,2004,104:832-839.
[172]Jia A, Zhang X H.cDNA cloning, characterization, and expression analysis of the Racl gene from Scophthalmus maximus[J]. Com Biochem PhyB,2009,154(1):80-84.
[173]Jantschko W, Georg F P, Zederbauer M, et al. Direct conversion of ferrous myeloperoxidase to compound Ⅱ by hydrogen peroxide:an anaerobic stopped-flow study[J]. Biochemical and Biophysical Research Communications,2003,312(2):292-298.
[174]Bradley P P, Priebat D A, Christensen R D, et al.Measurement of cutaneous inflammation estimation of neutrophil content with an enzyme marker [J]. Journal of Investigative Dermatology,1982,78(3): 206-209.
[175]Lieschke G J, Oates A C, Crowhurst M O, et al.Morp hologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish [J]. Blood,2001,98(10):3087-3096.
[176]Bermett C M, Kanki J P, Rhodes J, et al.Myelopoiesis in the zebrafish, Danio rerio[J].Blood,2001, 98(3):643-651.
[177]胡宝庆,刘毅,文春根.翘嘴鳜髓过氧化物酶基因克隆及重链序列的原核表达[J].山东大学学报(理学版),2009,44(5)：10-19.
[178]Yeh H Y, Klesius P H.Sequence analysis,charaeterization and tissue distribution of channelcatfish (Ictalurus punctatus Rafinesque,1818) myeloperoxidase cDNA[J].Fish shellfish Immunology,2010, 28(3):504-509.
[179]Johnson K R, Nauseef W M, Care A, et al. Characterization of cDNA clones for human myeloperoxidase: predicted amino acid sequence and evidence for multiple mRNA species [J].Nucleic Acids Research, 1987,15(5):2013-2028.
[180]Morishita K, Kubota N, Asano S, et al. Molecular cloning and characterization of cDNA for human myeloperoxidase[J]. The Journal of Biological Chemistry,1987,262(8):3844-3851.
[181]Zeng J, Fenna R E. X-ray crystal structure of canine myeloperoxidase at 3 A resolution[J]. Journal of Molecular Biology,1992,226(1):185-207.
[182]Furtmuller P G, Zederbaue M, Jantschko W, et al. Active site structure and catalytic mechanisms of human peroxidases[J]. Archives of Biochemistry and Biophysics,2006,445(2):199-213.
[183]朱元晓,赖春宁,王建安,等.人髓过氧化物酶基因cDNA克隆及急性白血病髓过氧化物酶基因的表达[J].生物工程学报,1993,9(1)：21-24.