黄鳝肠道cDNA文库构建、EST分析及抗菌肽基因的原核表达
|
摘要
黄鳝(Monopterus albus)是广泛分布于我国南北水域的名优水产养殖品种之一。本文运用分子生物学、生物信息学、基因工程等技术与方法,通过构建肠道cDNA文库对黄鳝肠道内营养代谢、免疫相关功能基因进行了一些初步分析与探讨；并对黄鳝抗菌肽hepcidin的原核表达进行了初步研究。
首次成功构建了黄鳝全肠未剪切cDNA文库。经检测,文库的库容量达到1.46×107cfu,重组阳性克隆率为96.88%,平均插入片段约2.11kb,符合高质量文库要求。随机从文库中挑取1056个阳性克隆进行5’单向测序,去除载体序列与低质量序列后获得了906条高质量的EST序列,平均长度为812bp,并提交到dbEST数据库(GenBank accession number:GW584265-GW585168)。通过拼接与组装,共获得762条有效的一致序列,其中61个重叠群包含2条以上的EST,其余701个仅含有一条EST。运用NCBI BLAST对序列进行比对,SeqGO进行功能注释。鉴定出与营养相关的EST共有5类,59种：蛋白质氨基酸代谢类25种,碳水化合物代谢类13种,脂类代谢类5种,能量代谢类12种,无机离子代谢类4种；发现免疫相关基因共73个,首次证实了肠道内存在大量与黄鳝免疫功能相关的基因,其中包括几丁质酶、溶菌酶、凝集素、抗氧化酶类、主要组织相容性复合体(MHC)、热休克蛋白(HSP)、IgM等；在黄鳝肠道中还发现了嗜酸性粒细胞趋化因子(ECC),这是该基因首次在硬骨鱼类中得到证实。
应用PCR技术从文库中筛选出了黄鳝抗菌肽基因hepcidin(GenBank accession number:GU997139),应用在线分析软件,对筛选的抗菌肽基因hepcidin进行了序列分析,发现其ORF区域为273bp,编码90个氨基酸残基的阳离子抗菌肽,分子量为10236Da。进一步分析抗菌肽蛋白质序列,包含24个氨基酸的信号肽和40个氨基酸的前域部分,成熟肽为26肽,结构预测显示其属于β-折叠类抗菌肽,包含有hepcidin类抗菌肽典型的8个半胱氨酸结构,形成4对二硫键。同源比对发现黄鳝hepcidin与多种动物的hepcidin序列高度同源,与大黄鱼和黑鲈hepcidin的相似性高达81%。
利用基因工程技术,首次对黄鳝hepcidin基因在原核细胞中表达进行了初步研究。分别构建了pET22b(+)重组表达载体,发现黄鳝抗菌肽hepcidin基因构建的pET22b(+)表达载体在原核细胞中不能表达。通过pET43. la(+)构建黄鳝hepcidin基因融合表达载体,转化到BL21(DE3)pLysS, BL21Star(DE3)pLysS和BL21-CondonPlus (DE3)-RIPL三种表达菌株中进行诱导表达,重组pET43. la(+)-hepcidin在原核细胞中均无明显表达。在pET32a(+)载体中分别构建黄鳝hepcidin全长编码序列(pET32a(+)-hepcidin)和去信号肽序列(pET32a(+)-nhepcidin)重组表达载体,转化到BL21(DE3)pLysS, BL21Star(DE3)pLysS, BL21-CondonPlus (DE3)-RIPL三种不同的表达菌株中进行诱导表达,发现含有信号肽的pET32a(+)-hepcidin重组表达载体在三种菌株中均不能正常表达；而不含信号肽的pET32a(+)-nhepcidin重组表达载体在BL21-CondonPlus (DE3)-RIPL中能高效表达。通过优化诱导表达条件,获得黄鳝抗菌肽pET32a(+)-nhepcidin重组载体在BL21-CondonPlus (DE3)-RIPL中的最佳表达条件为菌液浓度(OD600)0.5-0.7、温度为37℃、IPTG浓度0.5mM、诱导时间4-6h；鉴定发现pET32a (+)-nhepcidin在BL21-CodonPlus(DE3)-RIPL表达菌株中为可溶表达重组蛋白。
本试验首次成功构建了黄鳝肠道未剪切cDNA文库,发现了许多与肠道营养代谢和抗病相关的EST序列,为黄鳝肠道功能基因的研究奠定了分子基础；黄鳝肠道中抗菌肽hepcidin的发现不仅为黄鳝功能基因研究拓宽了道路,而且对开发用于黄鳝养殖的新型抗病药物提供了重要的候选物和参考依据。获得的研究资料将为后续研究打下基础。
The freshwater eel, Monopterus albus, is widely distributed throughout the world. In recent years it has become commercially important, resulting in the establishment of several facilities to culture this species in China. With the development of intensive aquaculture, several diseases, caused by bacteria, viruses and parasites, have emerged. We constructed an expressed sequence tag (EST) library (Uncut) to identify genes associated with nutrient metabolism and immunity in the intestine of M. albus, a commercially valuable species. Then preliminary study on M. albus hepcidin expression in prokaryotic expression vector.
Our objective was to identify nutrient metabolism and immunity-related genes in M. albus. We constructed a cDNA library from the intestinal tissue of M. albus. The library capacity reaches 1.46×107, the percentage of recombination is as high as 96.88%. PCR results showed that the average size of inserts was 2.11Kb. We sequenced 1056 randomly selected clones from the cDNA library. We obtained 906 high quality ESTs (GenBank accession number:GW584265-GW585168) with an average length 812 bp after trimming the vector sequence and eliminating low quality sequences. The ESTs were assembled into 61 contigs with an average length of 1057 bp, leaving 701 singletons. In total, we identified 762 unique sequences.Several (523 of 762) of the unique sequences had shared significant homology (BLASTX, score> 80, e-values<10-10) with known genes. We identified 59 species putative nutrition-related genes based on their functional classification in the Gene Ontology database. A number (73) of the unique sequences (including 8 contigs and 65 singlets) were related to immune function. We identified immunity-related genes that coded for chitinase, ferritin, MHC II, Hsp 70, HSP90, lectin, complement components, lysozyme, transferrin, glutathione peroxidase, serine protease inhibitor, and immunoglobulin. Eosinophil chemotactic cytokine (ECC) is a member of the chitinase family of proteins. Ours is the first report of this cytokine in teleosts.This study provides a basis for the development of the innate and adaptive immune systems in M. albus.
Application of PCR technology to screen out hepcidin (GenBank accession number: GU997139) antimicrobial peptide gene from the library. Bioinformatics analysis software, on the selection of the antimicrobial peptide hepcidin gene was sequenced, found that including a 273bp ORF region encoding 90 amino acid residues of the peptide. Further analysis of antimicrobial peptide protein sequence contains a signal peptide of 24 amino acids and pro-region part of 40 amino acids before the mature peptide of 26 amino acids, class of antimicrobial peptide hepcidin contains eight cysteine typical structure, the formation of four pairs of disulfide key; structure prediction shows that it belongs to P-sheet type peptide. Homologous than are found with a variety of animals, M. albus hepcidin sequences are highly homologous with the similarity of Larimichthys crocea croaker up to 81%.
Using genetic engineering techniques, preliminary study on M. albus hepcidin expression in prokaryotic expression vector. We constructed a direct expression vector pET22b (+) found that M. albus hepcidin peptide can not directly expression in pET22b (+), prove that this peptide with bactericidal activity. Then pET43.1a (+) construction of hepcidin fusion expression vector was transformed into BL21 (DE3) pLysS, BL21Star (DE3) pLysS and BL21-CondonPlus (DE3)-RIPL strain of three expression was induced and expressed recombinant protein had no significant expression. In pET32a (+) vector were constructed encoding full sequence of hepcidin (pET32a (+)-hepcidin) and no signal peptide sequence (pET32a (+)-nhepcidin) recombinant plasmid was transformed into BL21 (DE3) pLysS, BL21Star (DE3) pLysS, BL21-CondonPlus (DE3)-RIPL expression of three different strains induced for expression, found to contain a signal peptide of pET32a (+) -hepcidin fusion protein in three kinds of strains are not normal expression, while no signal peptide of pET32a (+)-nhepcidin recombinant protein can be highly expressed in BL21-CondonPlus (DE3)-RIPL. Induced by optimizing the conditions to obtain pET32a (+)-nhepcidin recombinant protein in BL21-CondonPlus (DE3)-RIPL in the best conditions for the expression of bacterial concentration (OD600) 0.5～0.7, temperature 37℃, IPTG concentration of 0.5mM, the induction time 4～6h. Identification found pET32a (+)-nhepcidin recombinant protein expressed in BL21-CodonPlus (DE3)-RIPL strain for soluble expression.
We successfully constructed cDNA library (uncut) from the intestinal tissue of M. albus and found a lot of nutrition metabolism and disease resistance-related EST sequences. Antimicrobial peptide hepcidin from the intestinal tissue was found not only functional gene of M. albus widening the road, but also the development of new disease for aquaculture provides an important candidate drugs and reference materials. Our study provides a basis for the identification of genetic variation among immune related genes and their relationship to disease resistance phenotypes.
首次成功构建了黄鳝全肠未剪切cDNA文库。经检测,文库的库容量达到1.46×107cfu,重组阳性克隆率为96.88%,平均插入片段约2.11kb,符合高质量文库要求。随机从文库中挑取1056个阳性克隆进行5’单向测序,去除载体序列与低质量序列后获得了906条高质量的EST序列,平均长度为812bp,并提交到dbEST数据库(GenBank accession number:GW584265-GW585168)。通过拼接与组装,共获得762条有效的一致序列,其中61个重叠群包含2条以上的EST,其余701个仅含有一条EST。运用NCBI BLAST对序列进行比对,SeqGO进行功能注释。鉴定出与营养相关的EST共有5类,59种：蛋白质氨基酸代谢类25种,碳水化合物代谢类13种,脂类代谢类5种,能量代谢类12种,无机离子代谢类4种；发现免疫相关基因共73个,首次证实了肠道内存在大量与黄鳝免疫功能相关的基因,其中包括几丁质酶、溶菌酶、凝集素、抗氧化酶类、主要组织相容性复合体(MHC)、热休克蛋白(HSP)、IgM等；在黄鳝肠道中还发现了嗜酸性粒细胞趋化因子(ECC),这是该基因首次在硬骨鱼类中得到证实。
应用PCR技术从文库中筛选出了黄鳝抗菌肽基因hepcidin(GenBank accession number:GU997139),应用在线分析软件,对筛选的抗菌肽基因hepcidin进行了序列分析,发现其ORF区域为273bp,编码90个氨基酸残基的阳离子抗菌肽,分子量为10236Da。进一步分析抗菌肽蛋白质序列,包含24个氨基酸的信号肽和40个氨基酸的前域部分,成熟肽为26肽,结构预测显示其属于β-折叠类抗菌肽,包含有hepcidin类抗菌肽典型的8个半胱氨酸结构,形成4对二硫键。同源比对发现黄鳝hepcidin与多种动物的hepcidin序列高度同源,与大黄鱼和黑鲈hepcidin的相似性高达81%。
利用基因工程技术,首次对黄鳝hepcidin基因在原核细胞中表达进行了初步研究。分别构建了pET22b(+)重组表达载体,发现黄鳝抗菌肽hepcidin基因构建的pET22b(+)表达载体在原核细胞中不能表达。通过pET43. la(+)构建黄鳝hepcidin基因融合表达载体,转化到BL21(DE3)pLysS, BL21Star(DE3)pLysS和BL21-CondonPlus (DE3)-RIPL三种表达菌株中进行诱导表达,重组pET43. la(+)-hepcidin在原核细胞中均无明显表达。在pET32a(+)载体中分别构建黄鳝hepcidin全长编码序列(pET32a(+)-hepcidin)和去信号肽序列(pET32a(+)-nhepcidin)重组表达载体,转化到BL21(DE3)pLysS, BL21Star(DE3)pLysS, BL21-CondonPlus (DE3)-RIPL三种不同的表达菌株中进行诱导表达,发现含有信号肽的pET32a(+)-hepcidin重组表达载体在三种菌株中均不能正常表达；而不含信号肽的pET32a(+)-nhepcidin重组表达载体在BL21-CondonPlus (DE3)-RIPL中能高效表达。通过优化诱导表达条件,获得黄鳝抗菌肽pET32a(+)-nhepcidin重组载体在BL21-CondonPlus (DE3)-RIPL中的最佳表达条件为菌液浓度(OD600)0.5-0.7、温度为37℃、IPTG浓度0.5mM、诱导时间4-6h；鉴定发现pET32a (+)-nhepcidin在BL21-CodonPlus(DE3)-RIPL表达菌株中为可溶表达重组蛋白。
本试验首次成功构建了黄鳝肠道未剪切cDNA文库,发现了许多与肠道营养代谢和抗病相关的EST序列,为黄鳝肠道功能基因的研究奠定了分子基础；黄鳝肠道中抗菌肽hepcidin的发现不仅为黄鳝功能基因研究拓宽了道路,而且对开发用于黄鳝养殖的新型抗病药物提供了重要的候选物和参考依据。获得的研究资料将为后续研究打下基础。
The freshwater eel, Monopterus albus, is widely distributed throughout the world. In recent years it has become commercially important, resulting in the establishment of several facilities to culture this species in China. With the development of intensive aquaculture, several diseases, caused by bacteria, viruses and parasites, have emerged. We constructed an expressed sequence tag (EST) library (Uncut) to identify genes associated with nutrient metabolism and immunity in the intestine of M. albus, a commercially valuable species. Then preliminary study on M. albus hepcidin expression in prokaryotic expression vector.
Our objective was to identify nutrient metabolism and immunity-related genes in M. albus. We constructed a cDNA library from the intestinal tissue of M. albus. The library capacity reaches 1.46×107, the percentage of recombination is as high as 96.88%. PCR results showed that the average size of inserts was 2.11Kb. We sequenced 1056 randomly selected clones from the cDNA library. We obtained 906 high quality ESTs (GenBank accession number:GW584265-GW585168) with an average length 812 bp after trimming the vector sequence and eliminating low quality sequences. The ESTs were assembled into 61 contigs with an average length of 1057 bp, leaving 701 singletons. In total, we identified 762 unique sequences.Several (523 of 762) of the unique sequences had shared significant homology (BLASTX, score> 80, e-values<10-10) with known genes. We identified 59 species putative nutrition-related genes based on their functional classification in the Gene Ontology database. A number (73) of the unique sequences (including 8 contigs and 65 singlets) were related to immune function. We identified immunity-related genes that coded for chitinase, ferritin, MHC II, Hsp 70, HSP90, lectin, complement components, lysozyme, transferrin, glutathione peroxidase, serine protease inhibitor, and immunoglobulin. Eosinophil chemotactic cytokine (ECC) is a member of the chitinase family of proteins. Ours is the first report of this cytokine in teleosts.This study provides a basis for the development of the innate and adaptive immune systems in M. albus.
Application of PCR technology to screen out hepcidin (GenBank accession number: GU997139) antimicrobial peptide gene from the library. Bioinformatics analysis software, on the selection of the antimicrobial peptide hepcidin gene was sequenced, found that including a 273bp ORF region encoding 90 amino acid residues of the peptide. Further analysis of antimicrobial peptide protein sequence contains a signal peptide of 24 amino acids and pro-region part of 40 amino acids before the mature peptide of 26 amino acids, class of antimicrobial peptide hepcidin contains eight cysteine typical structure, the formation of four pairs of disulfide key; structure prediction shows that it belongs to P-sheet type peptide. Homologous than are found with a variety of animals, M. albus hepcidin sequences are highly homologous with the similarity of Larimichthys crocea croaker up to 81%.
Using genetic engineering techniques, preliminary study on M. albus hepcidin expression in prokaryotic expression vector. We constructed a direct expression vector pET22b (+) found that M. albus hepcidin peptide can not directly expression in pET22b (+), prove that this peptide with bactericidal activity. Then pET43.1a (+) construction of hepcidin fusion expression vector was transformed into BL21 (DE3) pLysS, BL21Star (DE3) pLysS and BL21-CondonPlus (DE3)-RIPL strain of three expression was induced and expressed recombinant protein had no significant expression. In pET32a (+) vector were constructed encoding full sequence of hepcidin (pET32a (+)-hepcidin) and no signal peptide sequence (pET32a (+)-nhepcidin) recombinant plasmid was transformed into BL21 (DE3) pLysS, BL21Star (DE3) pLysS, BL21-CondonPlus (DE3)-RIPL expression of three different strains induced for expression, found to contain a signal peptide of pET32a (+) -hepcidin fusion protein in three kinds of strains are not normal expression, while no signal peptide of pET32a (+)-nhepcidin recombinant protein can be highly expressed in BL21-CondonPlus (DE3)-RIPL. Induced by optimizing the conditions to obtain pET32a (+)-nhepcidin recombinant protein in BL21-CondonPlus (DE3)-RIPL in the best conditions for the expression of bacterial concentration (OD600) 0.5～0.7, temperature 37℃, IPTG concentration of 0.5mM, the induction time 4～6h. Identification found pET32a (+)-nhepcidin recombinant protein expressed in BL21-CodonPlus (DE3)-RIPL strain for soluble expression.
We successfully constructed cDNA library (uncut) from the intestinal tissue of M. albus and found a lot of nutrition metabolism and disease resistance-related EST sequences. Antimicrobial peptide hepcidin from the intestinal tissue was found not only functional gene of M. albus widening the road, but also the development of new disease for aquaculture provides an important candidate drugs and reference materials. Our study provides a basis for the identification of genetic variation among immune related genes and their relationship to disease resistance phenotypes.
引文
Abu RS, Bloch SE, Lelkes PI, et al. Characterization of pardaxin-induced dopamine release from pheochromocytoma cells:role of calcium and eicosanoids[J]. J Pharmacol Exp Ther.1999;288(2):399～406.
Adams MD,Kelley JM, Gocayne JD, et al. Complementary DNA sequencing expressed sequence tags and the Human Genome Project[J]. Science, 1991,252(5013):1651-1656.
Aitken R, Gilchrist J, SinclairM C.Gene,1994;144(1):69-73.
Alexander JB, Ingram GA. Noncellular nonspecific defense mechanisms of fish[J]. Annu Rev Fish Dis,1992;2:249～79.
Altschul S F,Gish W,Miller W,et al. Basic local alignment search tool[J].Journal of Molecular Biology,1990;215:403-410.
Aoki T, Takano T, Santos MD, et al. Molecular innate immunity in teleost fish: review and future perspectives. In:Tsukamoto K, Kawamura T, Takeuchi T, Beard Jr TD, Kaiser MJ, editors. Fisheries for global welfare and environment.5th World Fisheries Congress;2008 p.263～76.
Apanius V, Penn D., Slev P R., et al.The nature of selection on the major histocompatibility complex[J]. Rev. Immunol,1999; 17(2):179～224.
Arai A, Naruse K, Mitani H, et al.Cloning and characterization of cDNAs for 70-kDa heat-shock proreins(Hsp70)from two fish species of the genus Oryzias[J]. Jpn J Genet,1995;70:423-433.
Aranishi F. Purification and characterization of alpha (1)-proteinase inhibitor from carp (Cyprinus carpio) serum[J]. Mar Biotechnol,1999;1:33～43.
Arason GJ. Lectins as defense molecules in vertebrates and invertebrates[J]. Fish and Shellfish Immunology 1996;6:277～289.
Ashburner M, Lewis SE. On ontologies for biologists:the Gene Ontology-uncouping the web.In:In Silico Biology.2002,Novartis Found Symp 247:66-80;discussion 80～83,84～90,244～252.
Balla G, Jacob HS, Balla J, et al. Ferritin:a cytoprotetive antioxidant stratagem of endothelium[J].J Biol Chem,1992;267(25):18148-18153.
Balla G, Jacob H S, Balla J, et al. Ferritin:a cytoprotetive antioxidant stratagem of endothelium[J].J Biol Chem,1992;267(25):18148-18153.
Bao B, Peatman E, Li P, et al. Catfish hepcidin gene is expressed in a wide range of tissues and exhibit tissues specific upregulation after bacterial infection[J]. Dev Comp Immunol,2005,29(11):939-950.
Bao B, Peatman E, Xu P, et al. The catfish liver-expressed antimicrobial peptide 2(LEAP-2)gene is expressed in a wide range of tissues and developmentally regulated[J]. Mol Immunol,2006,43(4):367～377.
Bayne CJ, Gerwick L. The acute phase response and innate immunity of fish[J]. Dev Comp Immunol,2001;25:725-43.
Bergljot, Magnadottir. Innate immunity of fish[J]. Fish and Shellfish Immunology, 2006;20:137-151.
Blake JA,Harris M.The Gene Ontology Project:Structured vocabularies for molecular biology and their application to genome and expression analysis.In:A.D.Baxevanis,D.B.Davison,R.Page,G.Stormo and L.Stein (eds),Current Protocols in Bioinformatics.2003,Wiley and Sona,Inc.,New York.
Bohana-Kashtan O., Ziporen L., Donin N., et al. Cell signals transduced by complemen[J]. Molecular Immunology,2004;41:583-97.
Bohley P., Seglen PO. Proteases and proteolysis in the lysosome[J]. Experientia, 1992;48:151-7.
Boot R G, Blommaart E F, Swart E, et al. Identification of a novel acidic mammalian chitinase distinct from chitotriosidase[J].J Biol Chem.2001,276(9):6770-6778
Bowden TJ, Butler R, Bricknell IR, et al. Serum trypsin-inhibitory activity in five species of farmed fish[J]. Fish and Shellfish Immunology,1997;7:377-85.
Campa-Cordova, A.I., Hernandez-Saavedra, N.Y., et al. Superoxide dismutase as modulator of immune function in American white shfimp (Litopenaeus vannamei) [J].Mol.Immunol,2002;44:943-9.
Carnevali O, Carletta R, Cambi A, et al. Yolk formation and degradation during oocyte maturation in seabream Sparus auratai:involvement of two lysosomal proteinases[J]. Biol Reprod,1999;60:140-6.
Carnevali O, Mosconi G, Cambi A, et al. Changes of lysosomal enzyme activities in sea bass (Dicentrarchus labrax) eggs and developing embryos[J]. Aquaculture,2001;202:249～56.
Chandran K, Sullivan NJ, Felbor U, et al. Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection[J]. Science,2005;308:1643～5.
Chen S L, Xu M Y, Ji X S, et al. Cloning, Characterization and expression analysis of hepcidin gene from red sea bream(chtysophrys major) [J]. Antimicro Agents Chemotherapy,2005;49(4):1608-1612.
Chen W, Fu X B., Ge S L., et al. Intravenous acid fibroblast growth factor protects intestinal mucosal cells against ischemia-reperfusion injury via regulating Bcl-2/Bax expression[J]. World J Gastroenterol,2005; 11(22):3419～3425.
Cheng Y, Zak O, Aisen P, et al. Structure of the human transferrin receptor-transferrin complex[J]. Cell,2004;116(4):565-576.
Cho JH. Cathepsin D produces antimicrobial peptide parasin 1 from histone H2A in the skin mucosa of fish[J].FASEB J,2002,16(3):429-431.
Chrast R, Verheijen MHG, Lemke G. Complement factors in adult peripheral nerve:a potential role in energy metabolism[J]. Neurochemistry International, 2004;45:353-9.
Conlon JM, Sower SA. Isolation of a peptide structurally related to mammalian corticostatins from the lamprey Petromyzon marinus[J].Comp Biochem Physiol,1996,114(2):133
Consuegra, S., C. Garcia de Leaniz. MHC-mediated mate choice increases parasite resistance in salmon[J]. Proc. R. Soc. B,2008;275:1397-1403.
Dabrowski K, Cieroszko A. Proteinase inhibitor(s) in seminal plasma of teleost fish[J]. J Fish Biol 1994;45:801-9.
Dalmo RA, Ingebrigsten K, Sveinbjornsson B, Seljelid R. Accumulation of immunomodulatory laminaran (beta (1,3)-D-glucan) in the heart, spleen and kidney of Atlantic cod, Gadus morhua[J]. J Fish Dis 1996;19:129～36.
Del Rio-Tsonis K, Tsonis PA, Zarkadis IK, et al. Expression of the third component of complement, C3, in regenerating limb blastema cells of urodeles[J]. Journal of Immunology,1998;161:6819～24.
Dong Z., Katar M., Linebaugh BE., et al. Expression of cathepsins B, D and L in mouse corneas infected with Pseudomonas aeruginosa[J]. Eur J Biochem 2001;268:6408～16.
Dorsehner M.O., Duris T., Bront C.R., et al. High levels of MHC class II allelic diversity in lake rainbow trout from Lake resevior[J].J Heredity, 2000;91(5):359～363.
Douglas S E, Gallant J W, Liebscher R S, et al. Identification and expression analysis of hepcidin21ike antimicrobial peptides in bony fish[J]. Dev Comp Immunol,2003;27:589-601.
Douglas S E,Gallant JW,Gong Z, et al. Cloning and developmental expression of a family of pleurocidin-like antimicrobial peptides from winter flounder,Pleuronecter americanus (walbaum) [J]. Dev Comp Immunol, 2001,25(2):137～147.
Ebert DH, Deussing J, Peters C, et al. Cathepsin L and cathepsin B mediate reovirus disassembly in murine fibroblast cells[J]. J Biol Chem 2002; 277:24609-17.
Ebran N,Julien S,Orange N,et al. Pore-forming properties and antibacterial activity of proteins extracted from epidermal mucus of fish[J]. Comp Biochem Physiol,1999,122(Part A):181～189.
Edwards SV, Hedrick PW. Evolution and ecology of MHC molecules:from genomics to sexual selection[J]. Trends Ecol Evol,1998;13(8):305～311.
Ellis AE. Inhibition of the Aeromonas salmonicida extracellular protease by a2-macroglobulin in the serum of rainbow trout[J]. Microb Pathog,1987;3:167-77.
Epple P, Apel K, Bohlmann H. ESTs reveal a multigene family for plant defensins in Arabidopsis thaliana[J]. FEBS Lett,1997,400(2):168-172.
Epsztejn S,Glickstein H, Picard V,et al. H-ferritin subunit overexpression in erythroid cells reduces the oxidative stress response and induces multidrug resistance properties[J]. Blood,1999;94 (10):3593～3603.
Epsztejn S, Glickstein H, Picard V, et al. H-ferritin subunit overexpression in erythroid cells reduces the oxidative stress response and induces multidrug resistance properties[J]. Blood,1999;94 (10):3593～3603.
Ewart KV, Johnson SC, Ross NW. Lectins of the innate immune system and their relevance to fish health[J]. ICES Journal of Marine Science,2001;58:380-5.
Feder M.E., Hofmann G.E. Heat-shock proteins, molecular chaperones, and the stress response:Evolutionary and ecological physiology[J]. Annual Review of Physiology,1999;61:243-282.
Fleming T.O., Klucken J., Katherine E. Small heat shock proteins protect against a-synuclein induced toxicity and aggregation[J]. Biochemical and Biophysical Research Communications,2006;351(3):631～638.
Flohe L. The selenoprotein glutathione peroxidase. In Glutathione-Chemical, Biochemical, and Medical Aspects, eds D Dolphin, O Avramovic, R Poulson, New York:Wiley-Interscience;1989,p.643～731.
Franz Villarroel, Adriana Bastias, Alin Casado, et al. Apolipoprotein A-I, an antimicrobial protein in Oncorhynchus mykiss:Evaluation of its expression in primary defence barriers and plasma levels in sick and healthy fish[J]. Fish & Shellfish Immunology,2007,23(1):197-209.
Ganassin RC.,Bols NC. Development of long-term rainbow trout spleen cultures that are haemopoietic and produce dendritic cells[J]. Fish and Shellfish Immunology 1996;6:17～34.
Ganz T. Hepcidin, a key regulator of iron metabolism and mediator of anemia of inflammation[J]. Blood,2003,102:783.
Germain, R. N. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation[J]. Cell,1994; 76:287.
Gocheva V, Zeng W, Ke D, et al. Distinct roles for cysteine cathepsin genes in multistage tumorigenesis[J]. Genes Dev,2006; 20:543-56.
Gonzalez R, Charlemagne J, Mahana W, et al. Specificity of natural serum antibodies present in phylogenetically distinct fish species[J]. Immunology 1988;63:31-36.
Gonzalez R, Matsiota P, Torchy C, et al. Natural anti-TNP antibodies from rainbow trout interfere with viral infection in vitro[J]. Research in Immunology,1989; 140:675-84.
Grimholt.U., Hordvik.I., Fosse V.M., et al. Molecular cloning of major histocompatibility complex class I cDNA from Atlantic salmon(salmon salar) [J]. Immunogenetics,1993;37(6):469-473.
Grinde B, Lie 0, Poppe T, Salte R. Species and individual variation in lysozyme activity in fish of interest in aquaculture[J]. Aquaculture,1988;68:299-304.
Grinde B. Lysozyme from rainbow trout, Salmo gairdneri Richardson, as an antibacterial agent against fish pathogens[J]. J Fish Dis,1989;12:95-104.
Gross PS, Bartllett TC, Browdy CI, et al. Immune gene discovery expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific white shrimp. Litopenaeus vannamei, and the Atlantic white shrimp, L.setiferus.Dev Comp Immunol,2001;25(7):565
Hairong Lu, Guodong Li, Hongyu Wu, et al. Fusion Expression of Antimicrobial Peptide GK1 in Escherichia coli[J]. Chin J Biotech,2008,24(1),21-26.
Hansen J D, Strassburger P, du Pasquier L.Conservation of an alpha-domain within the teleostean world, MHC class I from the rainbow trout, Oncorhynchus mykiss[J]. Dev Comp Immunol,1996;20(6):417-425.
Hara S, Yamakawa M. A novel antibacterial peptide family isolated from the silkworm, Bombyx mori[J].Biochem J,1995,310:651～655.
Hara S, Yamakawa M. Production in Escherichia coli of moricin, a novel type antibacterial peptide from the silkworm Bobyx mori[J]. Biochem Biophys Res Commun,1996,220:664-669.
Harder J, Barrels J, Christophers E, et al. Isolation and characterization of human beta-defensin-3,a novel human inducible peptide antibiotic[J]. J Biol Chem,2001,276(8):5707.
Hashimoto K., Nakanishi T., Kurosaw Y., et al. Isolation of carp genes encoding major histocompatibility complex antigens[J]. Proc Natl Acad Sci USA 1990; 87:6863-6867.
Hess CM, Edwards SV. The evolution of the major histocompatibility complex in birds[J]. BioScience,2002;52(5):423-31.
Hill D P, Blake JA, Richardson JE, et al. Extension and Integration of the Gene Ontology (GO):Combining GO vocabularies with external vocabularies [J]. Genome Res,2002,12:1982-1991.
Hiramatsu N, Ichikawa N, Fukada H, et al. Identification and characterization of proteases involved in specific proteolysis of vitellogenin and yolk proteins in salmonids [J]. J Exp Zool 2002;292:11～25.
Hirono I, Hwang J Y, Ono Y,et al. Two different types of hepcidins from the Japanese flounder Paralichtys olivaceus[J]. FEBS J,2005,272(20):5257～ 5264.
Hjelmeland K. Proteinase inhibitors in the muscle and serum of cod (Gadus morhua). Isolation and characterization[J]. Comp Biochem Physiol B, 1983;76:365-72.
Hochachka P.W., Somero G.N. Biochemical adaptation:mechanism and process in physiological evolution. Oxford University Press, New York. xi 2002, 466pp.
Hoffmann J A. Innate immunity of insects[J]. Curr Opin Immunol.1995;7:4～ 10.
Hordvik I, Grunholt U, Fosse V M, et al. Cloning and sequence analysis of cDNAs encoding the MHC class II beta chain in Atlantic salmon(Salmon salar) [J]. Immunogenetics 1993;37(6):437～441.
Hunter H N, Fulton D B, Ganz T,et al, the solution structure of human hepcidin,a peptide hormone with antimicrobial activity that is involved in iron uptake and hereditary hemochomatosis[J]. JBC,2002;277:238-250.
Hwang MD,Fung YW,Wang RX,et al.Analysis of expressed sequence tags from a fetal human heart cDNA library[J].Genomics,1995,30(2):293-298.
Ilyin G, Courselaud B, Troadec MB, et al. Comparative analysis of mouse hepcidin 1 and 2 genes:evidence for different patterns of expression and coinducibility during iron overlaod[J]. FEBS Lett,2003,542(1-3):22-26.
Ishida T, Zheng Z, Dichek Ice, et al. Molecular cloning of nonsecreted endothelial cell-derived lipase isofonns[J]. Genomics.2004 Jan,83(1):24～33
Jain C. Gene,1993; 133(1):99-102.
Jia HP, Schuae BC, Schudy A, et al. Discovery of new human β-defensins using a genomics base dapproach[J]. Gene,2001,263(122):211-218.
Jin FL, et al. Expression and characterization of antimicrobial peptide CecropinAD in the methylotrophic yeast Pichia pastoris[J]. Process Biochem (2008),doi:10.1016/j.procbio.2008.08.012
Jolles P, Jolles J. What's new in lysozyme research? Always a model system, today as yesterday[J]. Mol Cell Biochem 1984;63:165-89.
Kaattari SL, Piganelli JD. The specific immune response:humoral defense[M]//Iwama G, Nakanishi T. The fish immune system,organism,pathogen and environment.San Diego USA:Academic Press,1996:207～254.
Kachamakova NM, Irnazarow I, Parmentier HK, et al. Genetic differences in natural antibody levels in common carp (Cyprinus carpio L.) [J]. Fish and Shellfish Immunology 2006;21:404～13.
Katsumi Matsuzaki. Why and how are peptide-lipid interactions utilized for self-defense? Magainins and tachyplesins as archetypes 。 Biochimica et Biophysica Acta (BBA)-Biomembranes, Volume 1462, Issues 1-2,15 December 1999, Pages 1～10
Kimbrell D A, Bender B. The evolution and genetics of innate immunity[J].Natuer Rev.Genet.2001;2:256～267
Kimura Y., Madhavan M., Call MK., et al. Expression of complement 3 and complement 5 in newt limb and lens regeneration[J]. Journal of Immunology 2003;170:2331-9.
Kos J., Werle B., Lah T., Brunner N. Cysteine proteinases and their inhibitors in extracellular fluids:markers for diagnosis and prognosis in cancer[J]. Int J Biol Markers,2000; 15:84-89.
Kothary R.K., Jones. D., Candido E.P.70-kilodalton heat shock poly-peptides from rainbow trout:characterization of cDNA sequences[J]. Mol Cell Biol 1984;4:1785-1791.
Krause A, Neitz S, Magert HJ, et al. LEAP21, a novel highly disulfide bonded human peptide,exhibits antimicrobial activity[J]. FEBS Lett,2000;480:147～ 150.
Kulaksiz H, Theilig F, Bachmann S, et al. The iron-regulatory Peptide hormone hepcidin:expression and cellular localization in mammalian kidney[J] Journal of Endocrinolog,2005,184:361～370.
Lange S, Bambir S, Dodds AW, Magnado'ttir B. The ontogeny of complement component C3 in Atlantic cod (Gadus morhua L.)-an immunohistochemical study[J]. Fish and Shellfish Immunology,2004; 16:359～67.
Langston AL, Johnstone R, Ellis AE. The kinetics of the hypoferraemic response and changes in levels of alternative complement activity in diploid and triploid Atlantic salmon, following injection of lipopolysaccharide[J]. Fish and Shellfish Immunology,2001; 11:333～45.
Langston AL., Bricknell IR., Ellis AE. Iron binding capacity of peripheral blood leucocyte lysates from Atlantic salmon (Salmo salar L.). In:Barnes AC, Davidson GA, Hiney MP, McIntosh D, editors. Methodology in fish diseases research. Aberdeen:Fisheries Research Services,1998; p.111～6.
Lele Z, Engel S, Krone P.H. Hsp47 and hsp70 gene expression is differentially regulated in a stress and tissue-specific manner in zebrafish embryos[J]. Dev Genet 1997;21:123-133.
Lemaitre B.The road to Toll[J]. Nat Rev Immunol.2004;4(7):521～527.
Lie O, Evensen O, Sorensen A, Froysadal E. Study on lysozyme activity in some fish species[J]. Dis Aquat Org 1989;6:1～5.
Lindsay GJH., Gooday GW. Chitinolytic enzymes and the bacterial microflora in the digestive tract of cod, Gadus morhua[J]. J Fish Biol 1985;26:255～65.
Llanos RJ, Whitacre CM, Miceli DC. Potential involvement of C3 complement factor in amphibian fertilization[J]. Comparative Biochemistry and Physiology, Part A 2000;127:29～38.
Lomax J, McCray AT. Mapping the Gene Ontology into the Unified Medical Language System[J].Comp Funct Genomics,2004;5:345～361.
Lovoll M, Kilvik T, Boshra H, et al. Maternal transfer of complement components C3-1, C3-3, C3-4, C4, C5, C7, Bf, and Df to offspring in rainbow trout (Oncorhynchus mykiss) [J]. Immunogenetics,2006;58:168～79.
Magnadottir B, Gudmundsdottir S, Gudmundsdottir BK. Study of the humoral response of Atlantic salmon (Salmo salar L.), naturally infected with Aeromonas salmonicida ssp achromogenes[J]. Veterinary Immunology and Immunopathology,1995; 49:127～42.
Magnadottir B, Lange S, Steinarsson A, Gudmundsdottir S. The ontogenic development of innate immune parameters of cod (Gadus morhua L.) [J]. Comp Biochem Physiol B Biochem Mol Biol,2004; 139:217～24.
Manson FDC., Fletcher TC., Gooday GW. Localization of chitinolytic enzymes in blood of turbot, Scophthalmus maximus, and their possible roles in defense[J]. J Fish Biol,1992;40:919～27.
McGinnis S, Madden TL.BLAST:at the core of a powerful and diverse set of sequence analysis tools.Nucleic Acids Research,2004;32:W20-W25.
Melchedov S, de Ilarduya OM,Ohlrogge J.Towards a functional catalog of the plant genome:a survey of gene for lipid biosynthesis[J]. Plant Physiol, 2000;122(2):389-402.
Molina A, Biemar F, Muller F, et al.Cloning and expression analysis of an inducible HSP70 gene from tilapia fish[J].FEBS Lett,2000;474:5-10.
Moon JY, Henzler-Wildman KA, Ramamoorthy A. Expression and purification of a recombinant LL-37 from Escherichia coli[J]. Biochim Biophys Acta,2006;1758(9):1351-1358.
Morgan BP, Marchbank KJ, Longhi MP, et al. Complement:central to innate immunity and bridging to adaptive responses[J]. Immunological Letters, 2005;97:171-9.
Murphy JE, Robert C, Kupper TS. Interleukin-land cutaneous inflammation:a crucial link between innate and acquired immunity [J]. J Invest Dermatol,2000;114(3):602-608.
Mwthy TV. Expression of GST-fused kinase domain of human Csk homologous kinase from pichia pastoris facilitates easy purification[J]. Biotechnol Let,2004; 26(5):443-449.
Nam BH, Yamamoto E,Hirono I, et al. A survey of expressed genes in the leukocytes of Japanese founder, Paralichthy solivaceus, infected with Hiramerhabdovirus[J] Dev Comp Immunol,2000;24(1):13
Nemat 0 K, Saul R. The chitin catabolic cascade in the marine bacterium Vibrio fumissii[J]. JBiol Chem,1996;271(52):33425-33432.
Nemeth E, Rivera S, Gabayan V, et al. IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin[J]. J Clin Invest,2004; 113:1271.
Nicolas G, Chanvet C, Viatte L, et al. The gene encoding the iron regulatory peptide hepcidin is regulated by anemia, hypoxia, and inflammation[J]. The Journal of Clinical Investigation,2002,110,1037～1044.
Nicolas G, Bennoun M, Devaux I, et al. Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2)knockout mice[J]. Proc Natl Sci USA,2001;98:8780～8785.
Nicolas G, Bennoun M, Porteu A, et al. Severe iron deficiency amernia in transgenic mice expressing liver hepcidin[J]. Proceedings of the National Academy of Sciences of the United States of America,2002,99:4596-4601.
Noga E J,Fan Z, Silphaduang U. Histone-like proteins from fish are lethal to the parasitio dinoflagellate Amyloodinium ocellatum[J]. Parasitology,2001; 123(Ptl):57-65.
Nollen E.A.A., Morimoto R.I. Chaperoning signaling pathways:molecular chaperones as stress-sensing "heat shock" proteins. J Cell Sci 2002; 115:2809-2816. Takao S., Tetsuo K, Katsumasa G., et al.Possible role of calcineurin in heating-related increase of rat muscle mass. Biochemical and Biophysical Research Communications,2005;331(4):1301-1309.
Nonaka M, Smith SL. Complement system of bony and cartilaginous fish[J]. Fish and Shellfish Immunology,2000;10:215-28.
Oren Z, Shai Y. A class of highly potent antibacterial peptides derived from pardaxin:a pore-forming peptide isolated from moses sole fish Pardachirus marmoratus[J].r J Biochem,1996;237(1):303-310.
Owhashi M., Arita H., Hayai N. Identification of a novel eosinophil chemotactic cytokine (ECF-L) as a chitinase family protein[J]. J Biol Chem,2000; 275(2):1279-86.
P.C.特纳,A.G.迈克伦南,A.D.贝茨,M.R.H.怀特,等著；刘进元,李文君,王薛林等译校.分子生物学(第二版)[M]．北京：科学出版社,2004.
Parham P., Ohta T. Population biology of antigen presentation by MHC class I molecules[J]. Science,1996;272:67～74.
Park CH, Valore EV, Waring AJ, et al. Hepcidin, a urinary antimicrobial peptide synthesized in the liver[J]. Biol Chem,2001;276:7806-7810.
Park IY, Park CB, Kim MS, et al. Parasin I, an antimicrobial peptide derived from histone H2A in the catfish, Parasilurus asotus[J]. FEBS Lett,1998;437 (3):258～262.
Park K C, Osborne J A, Tsoi S C M, et al. Expressed sequence tags analysis of Atlantic halibut Hip poglossuship poglossus liver, kidney and spleen tissues following vaccination against Vibrio anguil larum and A eromonas salmonicida[J]. Fish Shellfish Immunol,2005;18:393～415.
Paul Y, Kassebaum C, Lazarovici P,et al. Translocation of acylated pardaxin into cells[J]. FFBS Letters,1998;440(122):131-134.
Peng L, Xu Z, Fang X, et al. High—level expression of soluble human β-defensin-2 in Escherichia coli[J]. Process Biochem,2004,39(12):2199.
Persson A.C., Stet.R.JM., Pilstrom L., et al. Characterization of MHC class 1 and beta2-microglobulin sequences in Atlantic cod revealing an unusual high number of expressed class I genes[J].Immunogenetics,1999;50(1):49～59.
Pigeon C, Ilyin G, Courselaud B, et al. A new mouse liver-specific gene, encoding a protein homologous to human antimicrobial peptide hepcidin, is overexpressed during iron overload[S].J Biol Chem,2001,276(11):7811～ 7819.
Pouny Y, Rapaport D, et al. Interaction of antimicrobial dermaseptin and its fluorescently labeled analogues with phospholipid membranes [J]. Biochemistry,1992,31(49):12416-12423.
Press MCL, Dannevig BH, Landsverk T. Immune and enzyme histochemical phenotypes of lymphoid and nonlymphoid cells within the spleen and head kidney of Atlantic salmon (Salmo salar L.) [J]. Fish and Shellfish Immunology,1994;4:79-93.
Qiu Z, Hingley ST, Simmons G, et al. Endosomal proteolysis by cathepsins is necessary for murine coronavirus mouse hepatitis virus type 2 spike-mediated entry[J]. J Virol,2006;80:5768-76.
Raldu'a D, Fabra M, Bozzo MG, et al. Cathepsin B-mediated yolk protein degradation during killifish oocyte maturation is blocked by an HATPase inhibitor:effects on the hydration mechanism[J]. Am J Physiol Regul Integr Comp Physiol,2006;290:456～466.
Rao XC, Li S, Hu JC, et al. A novel carrier molecule for high-level expression of peptide antibiotics in Escherichia coli[J]. Pro Exp Purif,2004,36:11～18.
Robertsen B. The interferon system of teleost fish[J]. Fish and Shellfish Immunology,2006;20(2):172-191.
Roeder T. Solid-phase cDNA library construction, a versatile apporach[J]. Nucleic Acids Res,1998,26(14):3451-3452.
Russell S., Lumsden JS. Function and heterogeneity of fish lectins[J]. Veterinary Immunology and Immunopathology,2005; 108:111～120.
Salte R, Gjoen HM, Norberg K, et al. Plasma protein levels as potential marker traits for resistance to furunculosis[J]. J Fish Dis,1993;16:561～8.
Sebastian P, Wallwits J, Schmidt S. Semi automated production of a set of different recombinant GST-Streptag fusion protein[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2003,786(1-2):343～355.
Shai Y. Mechanism of the binding, insertion and destabilization of phospholipid bilayer membranes by a-helical antimicrobial and cell non-selective membrane-lytic peptides[J].Biochim Biophys Acta,1999;1462:55～70.
Shai Y. Pardaxin:channel formation by a shark repellant peptide from fish[J]. Toxicology,1994,87(13):109～129.
Sharmeks A D. Gene,1994; 138(1-2):105-108.
Sheridan M A. Lipid dynamics in fish:aspects of absorption, transportation, deposition and mobilization[J]. Comp. Biochem. Physiol,1988,90(4): 670～690.
Sherwin T, Brookes NH, Loh IP, Poole CA, Clover GM. Cellular incursion into Bowman's membrane in the peripheral cone of the keratoconic cornea[J]. Exp Eye Res 2002;74:473～482.
Shike H, Lauth X, Westerman M E, et al. Bass hepcidin is a novel antimicrobial peptide induced by bacterial challenge[J]. Eur J Biochem,2002,269(8): 2232～2237.
Shike H, Shimizu C, Lauth X, et al. Organization and expression analysis of the zebrafish hepcidin gene, antimicrobial peptide gene conserved among vertebrates[J]. Dev Comp Immunol,2004,28:747～754.
Shona K, Whyte. The innate immune response of finfish-A review of current knowledge[J]. Fish and Shellfish Immunology,2007;23:1127-1151.
Silphaduang U, Edwardb JN. Peptide antibiotics in mast cells of fish[J].Nature,2001,414(6861):268-269.
Sommer S. The importance of immune gene variability (MHC) in evolutionary ecology and conservation[J]. Front. Zool.2005;2:1～18.
Song LC, Xiang SJ, Guo CY, et al. Cloning, characterization, and expression analysis of hepcidin gene from Red Sea bream(Chrysophrys major)[J]. AAC, 2005,49(4):1608-1612.
Stafford JL, Belosevic M. Transferrin and the innate immune response of fish: identification of a novel mechanism of macrophage activation[J]. Dev Comp Immunol,2003;27:539-554.
Stafford JL, Neumann NF, Belosevic M. Products of proteolytic cleavage of transferrin induce nitric oxide response of goldfish macrophages[J]. Dev Comp Immunol,2001;25:101-15.
Sugano S, Maruyama K. Toward the construction of a full length cDNA library[J]. Tanpakushitsu Kakusan Koso,1993,38:476～484.
Sultmann H., Mayer W.E., Mayer F. Organization of MHC class II B genes in the zebra fish(Brachydanio reio) [J].Genomics 1994;23(1):1～4.
Susan E D, et al. Identification and expression analysis of hepcidin-like antimicrobial peptide in bony fish [J]. Developmental and Comparative Immunology,2003,27:589～601.
Suzuki M,Morimatsu M, Yamashita T,et al. A novel serum chitinase that is expressed in bovine liver[J]. FEBS Lett.2001;506(2):127-130.
Takeuchi H, Figueroa F, O'hUigin C, et al.Cloning and characterization of class I Mhc genes of the zebrafish, Danio rerio[J]. Immunogenetics,1995;42(2):77～ 84.
Tateno H, Saneyoshi A, Ogawa T, et al. Isolation and characterization of rhamnose-binding lectins from eggs of steelhead trout (Oncorhynchus mykiss) homologous to low density lipoprotein receptor superfamily[J]. Journal of Biological Chemistry 1998;273(30):19190-7.
Tort L, Balasch JC, Mackenzie S. Fish immune system. A crossroads between innate and adaptive responses[J]. Immunologi'a 2003;22(3):277-86.
Townes CL, Michailidis G, Nile CJ, et al. Induction of cationic chicken liver-expressed antimicrobial peptide 2 in response to Salmonella enterica infection[J]. Infect Immun,2004,72(12):6987～6993
Townsend, A., and H. Bodmer. Antigen recognition by class I-restricted T lymphocytes[J]. Annu. Rev. Immunol,1989;7:601.
Vanerp S H M., Dixon B, Figneroa F, et al. Identification and characterization of new major histocompatibility complex class I gene in carp (Cyrinus carpio) [J]. Immunogenetics,1996a;44(1):49-61.
Vanerp S.H.M., Egberts E., Stet R.J. Characterization of major histocompatibility complex class ⅡA and B genes in gynogenetic carp clone[J]. Immunogenetics, 1996b,44(3):192～202.
Vilain C, Wetzel M-C, Du Pasuier L, et al. Structural and functional analysis of spontaneous anti-nitrophenyl antibodies in three cyprinid fish species:carp (Cyprinus carpio), goldfish (Carassius auratus) and tench (Tinca tinca) [J]. Developmental and Comparative Immunology,1984;8:611-22.
Wang RF, Wang X, Johnston SL, et al. Development of a retrovirus-based complementary DNA expression system for the cloning of tumor antigens[J].Cancer Res,1998,58 (16):3519-3525.
Wang Z, Zhang S, Wang G, et al. Complement activity in the egg cytosol of zebrafish Danio rerio:evidence for the defense role of maternal complement components[J]. Plos One,2008;3(1):1463
Weinstein D A, Roy C N, Fleming M D,et al. Inappropriate expression of hepcidin is associated with iron refractory anemia:Implications for the anemia of chronic disease[J]. Blood,2002,100:37-76.
Yang M, Wang KJ, Zhou HL. A novel hepcidin-like antimicrobial peptide isolated from the Japanese Bass[J]. J Xiamen University (Natural Science),2004, 43(3):286～287.
Yano T. The nonspecific immune system:humoral defense. In:Iwama G, Nakanishi T, editors. The fish immune system:organism, pathogen, and environment. San Diego, USA:Academic Press;1996,p.105-53.
Yeaman MR, Yount NY. Mechanisms of antimicrobial peptide action and resistanc[J]. Pharmacol.Rev.2003;55 (1):27～55
Yifeng Li, Xia Li, Guangshun Wang.Cloning, expression, isotope labeling, and purification of human antimicrobial peptide LL-37 in Escherichia coli for NMR studies[J]. Protein Expression and Purification,2006,47:498-505.
Yousif AN., Albright LJ., Evelyn TPT. In vitro evidence for the antibacterial role of lysozyme in salmonid eggs[J]. Dis Aquat Org,1994;19:15-9.
Zasloff. Antimicrobial peptides of multicellular organisms[J]. Nature, 2002;415:389-395.
Zhang L, Falls T, Wu M, et al.Determinants of recombinant productionof antimicrobial cationic peptides and creation of peptide variant inbacteria[J].Biochem Biophys. Res. Commun.1998,247:674～680.
Zhang Y A, Zou J, Chang C I, et al. Discovery and characterization of two types of liver-expressed antimicrobial peptide 2(LEAP-2)genes in rainbow trout[J]. Vet Immunol Immunopathol,2004,101:259～269.
Zhao, D X, Song S H, Wang Q, et al., Discovery of immune-related genes in Chinese mitten crab (Eriocheir sinensis) by expressed sequence tag analysis of haemocytes[J]. Aquaculture,2009,283 (3-4):297-303.
Zugel U, Kaufmann S.H.E. Role of Heat Shock Proteins in Protection from and Patho genesis of Infectious Diseases. Clin. Microbiol. Rev,1999; 12:19～ 39.
Zuo X, Woo PTK. In vivo neutralization of Cryptobia salmositica metallo-protease by alpha 2-macroglobulin in the blood of rainbow trout Oncorhynchus mykiss and in brook charr Salvelinus fontinalis[J]. Dis Aquat Org, 1997;29:67-72.
陈文波,刘黎黎,李文笙,等.鲤鱼肝胰脏cDNA文库的表达序列标签分析及Lb-Fabp mRNA的特征[J].水生生物学报,2009,33(3)：376～384。
方永强,中庆祥,翁幼竹,等.文昌鱼头部cDNA文库的构建[J].台湾海峡,2003,22(2)：131-135.
胡国良,曹华斌,郭小权,等.高能低蛋白日粮对蛋鸡肝载脂蛋白A-I基因mRNA表达的影响[J].中国兽医科学2009,39(7)：641-645.
胡松年主编.基因表达序列标签(EST)数据分析手册[M].杭州：浙江大学出版社,2005.
姜秀云.牛分枝杆菌主要保护性抗原基因的克隆、表达及免疫研究[D].吉林农业大学博士论文,2005.
赖玉平,彭沂非,郁正艳,等.DCD-1L在毕赤酵母中的克隆和表达[J].中国生物工程杂志.2004,24(2)：61-65.
李伟,陈松林.牙鲆抗菌肽hepcidin基因在大肠杆菌中的融合表达[J].高技术通讯.2007,17(7)：765-770.
李旭霞,逢越,肖蓉,等.日本七鳃鳗(Lampetrs japonica)唾液腺cDNA文库的构建[J].辽宁师范大学学报(自然科学版),2004,27(1)：73～75.
李志琼,余冰,张克英,等.饲粮中添加a-亚麻酸对产蛋鸡载脂蛋白B和血脂的影响[J].动物营养学报,2008,20(6)：674-678.
刘家芳,杨代勤,阮国良.黄鳝消化酶活性的研究[J].湖北农业科学,2007,46(5)：804-807.
彭彩云.黄鳝养殖常见病害的防治[J].现代农业科技,2009,14：304～305
申艳敏,魏建超,尚书文等.人源抗菌肽LL-37在毕赤酵母中的高效表达及其活性检测[J].微生物学通报.2008,35(4)：539～544.
唐勇.黄鳝抗菌活性物质的制备及其抑菌效果的研究[D].四川农业大学硕士论文,2007.
王克坚,周红玲,杨明.海水养殖鲈鱼分离出一种hepcidin抗菌肽新基因[J].厦门大学学报：自然科学版,2004,43(3)：286～287.
吴乃虎.基因工程原理(第二版)[M].北京：科学出版社,1999.
徐奇友,单安山,王安.乳铁蛋白对早期断奶仔猪组织中微量元素含量的影响 [J].动物营养学报,2005,17(4)：62.
杨芳,贺智敏,詹显全,等.大鲵肝脏组织定向cDNA文库的构建及鉴定[J].动物学报,2004,50(3)：475-478.
杨贵贞等著.免疫活性细胞的效应[M].北京：科学出版社,1986
杨明,王克坚,周红玲,等.我国海水养殖真鲷分离出hepcidin抗菌肽新基因[J].高技术通讯,2004,14(增刊)：323～328.
杨旭.抗菌肽Pexiganan和IB-367基因在大肠杆菌中的克隆与表达[D].天津大学硕士论文.2006
药立波,常智杰,马文丽.医学分子生物学实验技术[J].北京：人民卫生出版社,2002.164.
尹娜,李鸿钧,彭梅等.抗菌肽Cecropin D在毕赤酵母中的表达、纯化及活性鉴定[J].中国生物制品学杂志,2008,21(3)：185～189.
张勇,张为民,李欣,等.石斑鱼卵巢cDNA文库构建及脂肪酸结合蛋白克隆[J].中山大学学报(自然科学版),2003,42(2)：66-69.
张宁.大海马cDNA文库构建、分析及抗菌肽新基因的初步功能研究[D],中山大学博士论文,2007.
张学俊,屈刚,朱文漓,等.草鱼肠道cDNA文库构建及部分ESTs分析[J].水生生物学报,2007,31(2)：251-258.
张雅丽,杨国庆,郭英,陈永福.密码子优化的牛精蛋白基因在大肠杆菌中的表达[J].高技术通讯,2002，4(1)：42～46.
Adams MD,Kelley JM, Gocayne JD, et al. Complementary DNA sequencing expressed sequence tags and the Human Genome Project[J]. Science, 1991,252(5013):1651-1656.
Aitken R, Gilchrist J, SinclairM C.Gene,1994;144(1):69-73.
Alexander JB, Ingram GA. Noncellular nonspecific defense mechanisms of fish[J]. Annu Rev Fish Dis,1992;2:249～79.
Altschul S F,Gish W,Miller W,et al. Basic local alignment search tool[J].Journal of Molecular Biology,1990;215:403-410.
Aoki T, Takano T, Santos MD, et al. Molecular innate immunity in teleost fish: review and future perspectives. In:Tsukamoto K, Kawamura T, Takeuchi T, Beard Jr TD, Kaiser MJ, editors. Fisheries for global welfare and environment.5th World Fisheries Congress;2008 p.263～76.
Apanius V, Penn D., Slev P R., et al.The nature of selection on the major histocompatibility complex[J]. Rev. Immunol,1999; 17(2):179～224.
Arai A, Naruse K, Mitani H, et al.Cloning and characterization of cDNAs for 70-kDa heat-shock proreins(Hsp70)from two fish species of the genus Oryzias[J]. Jpn J Genet,1995;70:423-433.
Aranishi F. Purification and characterization of alpha (1)-proteinase inhibitor from carp (Cyprinus carpio) serum[J]. Mar Biotechnol,1999;1:33～43.
Arason GJ. Lectins as defense molecules in vertebrates and invertebrates[J]. Fish and Shellfish Immunology 1996;6:277～289.
Ashburner M, Lewis SE. On ontologies for biologists:the Gene Ontology-uncouping the web.In:In Silico Biology.2002,Novartis Found Symp 247:66-80;discussion 80～83,84～90,244～252.
Balla G, Jacob HS, Balla J, et al. Ferritin:a cytoprotetive antioxidant stratagem of endothelium[J].J Biol Chem,1992;267(25):18148-18153.
Balla G, Jacob H S, Balla J, et al. Ferritin:a cytoprotetive antioxidant stratagem of endothelium[J].J Biol Chem,1992;267(25):18148-18153.
Bao B, Peatman E, Li P, et al. Catfish hepcidin gene is expressed in a wide range of tissues and exhibit tissues specific upregulation after bacterial infection[J]. Dev Comp Immunol,2005,29(11):939-950.
Bao B, Peatman E, Xu P, et al. The catfish liver-expressed antimicrobial peptide 2(LEAP-2)gene is expressed in a wide range of tissues and developmentally regulated[J]. Mol Immunol,2006,43(4):367～377.
Bayne CJ, Gerwick L. The acute phase response and innate immunity of fish[J]. Dev Comp Immunol,2001;25:725-43.
Bergljot, Magnadottir. Innate immunity of fish[J]. Fish and Shellfish Immunology, 2006;20:137-151.
Blake JA,Harris M.The Gene Ontology Project:Structured vocabularies for molecular biology and their application to genome and expression analysis.In:A.D.Baxevanis,D.B.Davison,R.Page,G.Stormo and L.Stein (eds),Current Protocols in Bioinformatics.2003,Wiley and Sona,Inc.,New York.
Bohana-Kashtan O., Ziporen L., Donin N., et al. Cell signals transduced by complemen[J]. Molecular Immunology,2004;41:583-97.
Bohley P., Seglen PO. Proteases and proteolysis in the lysosome[J]. Experientia, 1992;48:151-7.
Boot R G, Blommaart E F, Swart E, et al. Identification of a novel acidic mammalian chitinase distinct from chitotriosidase[J].J Biol Chem.2001,276(9):6770-6778
Bowden TJ, Butler R, Bricknell IR, et al. Serum trypsin-inhibitory activity in five species of farmed fish[J]. Fish and Shellfish Immunology,1997;7:377-85.
Campa-Cordova, A.I., Hernandez-Saavedra, N.Y., et al. Superoxide dismutase as modulator of immune function in American white shfimp (Litopenaeus vannamei) [J].Mol.Immunol,2002;44:943-9.
Carnevali O, Carletta R, Cambi A, et al. Yolk formation and degradation during oocyte maturation in seabream Sparus auratai:involvement of two lysosomal proteinases[J]. Biol Reprod,1999;60:140-6.
Carnevali O, Mosconi G, Cambi A, et al. Changes of lysosomal enzyme activities in sea bass (Dicentrarchus labrax) eggs and developing embryos[J]. Aquaculture,2001;202:249～56.
Chandran K, Sullivan NJ, Felbor U, et al. Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection[J]. Science,2005;308:1643～5.
Chen S L, Xu M Y, Ji X S, et al. Cloning, Characterization and expression analysis of hepcidin gene from red sea bream(chtysophrys major) [J]. Antimicro Agents Chemotherapy,2005;49(4):1608-1612.
Chen W, Fu X B., Ge S L., et al. Intravenous acid fibroblast growth factor protects intestinal mucosal cells against ischemia-reperfusion injury via regulating Bcl-2/Bax expression[J]. World J Gastroenterol,2005; 11(22):3419～3425.
Cheng Y, Zak O, Aisen P, et al. Structure of the human transferrin receptor-transferrin complex[J]. Cell,2004;116(4):565-576.
Cho JH. Cathepsin D produces antimicrobial peptide parasin 1 from histone H2A in the skin mucosa of fish[J].FASEB J,2002,16(3):429-431.
Chrast R, Verheijen MHG, Lemke G. Complement factors in adult peripheral nerve:a potential role in energy metabolism[J]. Neurochemistry International, 2004;45:353-9.
Conlon JM, Sower SA. Isolation of a peptide structurally related to mammalian corticostatins from the lamprey Petromyzon marinus[J].Comp Biochem Physiol,1996,114(2):133
Consuegra, S., C. Garcia de Leaniz. MHC-mediated mate choice increases parasite resistance in salmon[J]. Proc. R. Soc. B,2008;275:1397-1403.
Dabrowski K, Cieroszko A. Proteinase inhibitor(s) in seminal plasma of teleost fish[J]. J Fish Biol 1994;45:801-9.
Dalmo RA, Ingebrigsten K, Sveinbjornsson B, Seljelid R. Accumulation of immunomodulatory laminaran (beta (1,3)-D-glucan) in the heart, spleen and kidney of Atlantic cod, Gadus morhua[J]. J Fish Dis 1996;19:129～36.
Del Rio-Tsonis K, Tsonis PA, Zarkadis IK, et al. Expression of the third component of complement, C3, in regenerating limb blastema cells of urodeles[J]. Journal of Immunology,1998;161:6819～24.
Dong Z., Katar M., Linebaugh BE., et al. Expression of cathepsins B, D and L in mouse corneas infected with Pseudomonas aeruginosa[J]. Eur J Biochem 2001;268:6408～16.
Dorsehner M.O., Duris T., Bront C.R., et al. High levels of MHC class II allelic diversity in lake rainbow trout from Lake resevior[J].J Heredity, 2000;91(5):359～363.
Douglas S E, Gallant J W, Liebscher R S, et al. Identification and expression analysis of hepcidin21ike antimicrobial peptides in bony fish[J]. Dev Comp Immunol,2003;27:589-601.
Douglas S E,Gallant JW,Gong Z, et al. Cloning and developmental expression of a family of pleurocidin-like antimicrobial peptides from winter flounder,Pleuronecter americanus (walbaum) [J]. Dev Comp Immunol, 2001,25(2):137～147.
Ebert DH, Deussing J, Peters C, et al. Cathepsin L and cathepsin B mediate reovirus disassembly in murine fibroblast cells[J]. J Biol Chem 2002; 277:24609-17.
Ebran N,Julien S,Orange N,et al. Pore-forming properties and antibacterial activity of proteins extracted from epidermal mucus of fish[J]. Comp Biochem Physiol,1999,122(Part A):181～189.
Edwards SV, Hedrick PW. Evolution and ecology of MHC molecules:from genomics to sexual selection[J]. Trends Ecol Evol,1998;13(8):305～311.
Ellis AE. Inhibition of the Aeromonas salmonicida extracellular protease by a2-macroglobulin in the serum of rainbow trout[J]. Microb Pathog,1987;3:167-77.
Epple P, Apel K, Bohlmann H. ESTs reveal a multigene family for plant defensins in Arabidopsis thaliana[J]. FEBS Lett,1997,400(2):168-172.
Epsztejn S,Glickstein H, Picard V,et al. H-ferritin subunit overexpression in erythroid cells reduces the oxidative stress response and induces multidrug resistance properties[J]. Blood,1999;94 (10):3593～3603.
Epsztejn S, Glickstein H, Picard V, et al. H-ferritin subunit overexpression in erythroid cells reduces the oxidative stress response and induces multidrug resistance properties[J]. Blood,1999;94 (10):3593～3603.
Ewart KV, Johnson SC, Ross NW. Lectins of the innate immune system and their relevance to fish health[J]. ICES Journal of Marine Science,2001;58:380-5.
Feder M.E., Hofmann G.E. Heat-shock proteins, molecular chaperones, and the stress response:Evolutionary and ecological physiology[J]. Annual Review of Physiology,1999;61:243-282.
Fleming T.O., Klucken J., Katherine E. Small heat shock proteins protect against a-synuclein induced toxicity and aggregation[J]. Biochemical and Biophysical Research Communications,2006;351(3):631～638.
Flohe L. The selenoprotein glutathione peroxidase. In Glutathione-Chemical, Biochemical, and Medical Aspects, eds D Dolphin, O Avramovic, R Poulson, New York:Wiley-Interscience;1989,p.643～731.
Franz Villarroel, Adriana Bastias, Alin Casado, et al. Apolipoprotein A-I, an antimicrobial protein in Oncorhynchus mykiss:Evaluation of its expression in primary defence barriers and plasma levels in sick and healthy fish[J]. Fish & Shellfish Immunology,2007,23(1):197-209.
Ganassin RC.,Bols NC. Development of long-term rainbow trout spleen cultures that are haemopoietic and produce dendritic cells[J]. Fish and Shellfish Immunology 1996;6:17～34.
Ganz T. Hepcidin, a key regulator of iron metabolism and mediator of anemia of inflammation[J]. Blood,2003,102:783.
Germain, R. N. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation[J]. Cell,1994; 76:287.
Gocheva V, Zeng W, Ke D, et al. Distinct roles for cysteine cathepsin genes in multistage tumorigenesis[J]. Genes Dev,2006; 20:543-56.
Gonzalez R, Charlemagne J, Mahana W, et al. Specificity of natural serum antibodies present in phylogenetically distinct fish species[J]. Immunology 1988;63:31-36.
Gonzalez R, Matsiota P, Torchy C, et al. Natural anti-TNP antibodies from rainbow trout interfere with viral infection in vitro[J]. Research in Immunology,1989; 140:675-84.
Grimholt.U., Hordvik.I., Fosse V.M., et al. Molecular cloning of major histocompatibility complex class I cDNA from Atlantic salmon(salmon salar) [J]. Immunogenetics,1993;37(6):469-473.
Grinde B, Lie 0, Poppe T, Salte R. Species and individual variation in lysozyme activity in fish of interest in aquaculture[J]. Aquaculture,1988;68:299-304.
Grinde B. Lysozyme from rainbow trout, Salmo gairdneri Richardson, as an antibacterial agent against fish pathogens[J]. J Fish Dis,1989;12:95-104.
Gross PS, Bartllett TC, Browdy CI, et al. Immune gene discovery expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific white shrimp. Litopenaeus vannamei, and the Atlantic white shrimp, L.setiferus.Dev Comp Immunol,2001;25(7):565
Hairong Lu, Guodong Li, Hongyu Wu, et al. Fusion Expression of Antimicrobial Peptide GK1 in Escherichia coli[J]. Chin J Biotech,2008,24(1),21-26.
Hansen J D, Strassburger P, du Pasquier L.Conservation of an alpha-domain within the teleostean world, MHC class I from the rainbow trout, Oncorhynchus mykiss[J]. Dev Comp Immunol,1996;20(6):417-425.
Hara S, Yamakawa M. A novel antibacterial peptide family isolated from the silkworm, Bombyx mori[J].Biochem J,1995,310:651～655.
Hara S, Yamakawa M. Production in Escherichia coli of moricin, a novel type antibacterial peptide from the silkworm Bobyx mori[J]. Biochem Biophys Res Commun,1996,220:664-669.
Harder J, Barrels J, Christophers E, et al. Isolation and characterization of human beta-defensin-3,a novel human inducible peptide antibiotic[J]. J Biol Chem,2001,276(8):5707.
Hashimoto K., Nakanishi T., Kurosaw Y., et al. Isolation of carp genes encoding major histocompatibility complex antigens[J]. Proc Natl Acad Sci USA 1990; 87:6863-6867.
Hess CM, Edwards SV. The evolution of the major histocompatibility complex in birds[J]. BioScience,2002;52(5):423-31.
Hill D P, Blake JA, Richardson JE, et al. Extension and Integration of the Gene Ontology (GO):Combining GO vocabularies with external vocabularies [J]. Genome Res,2002,12:1982-1991.
Hiramatsu N, Ichikawa N, Fukada H, et al. Identification and characterization of proteases involved in specific proteolysis of vitellogenin and yolk proteins in salmonids [J]. J Exp Zool 2002;292:11～25.
Hirono I, Hwang J Y, Ono Y,et al. Two different types of hepcidins from the Japanese flounder Paralichtys olivaceus[J]. FEBS J,2005,272(20):5257～ 5264.
Hjelmeland K. Proteinase inhibitors in the muscle and serum of cod (Gadus morhua). Isolation and characterization[J]. Comp Biochem Physiol B, 1983;76:365-72.
Hochachka P.W., Somero G.N. Biochemical adaptation:mechanism and process in physiological evolution. Oxford University Press, New York. xi 2002, 466pp.
Hoffmann J A. Innate immunity of insects[J]. Curr Opin Immunol.1995;7:4～ 10.
Hordvik I, Grunholt U, Fosse V M, et al. Cloning and sequence analysis of cDNAs encoding the MHC class II beta chain in Atlantic salmon(Salmon salar) [J]. Immunogenetics 1993;37(6):437～441.
Hunter H N, Fulton D B, Ganz T,et al, the solution structure of human hepcidin,a peptide hormone with antimicrobial activity that is involved in iron uptake and hereditary hemochomatosis[J]. JBC,2002;277:238-250.
Hwang MD,Fung YW,Wang RX,et al.Analysis of expressed sequence tags from a fetal human heart cDNA library[J].Genomics,1995,30(2):293-298.
Ilyin G, Courselaud B, Troadec MB, et al. Comparative analysis of mouse hepcidin 1 and 2 genes:evidence for different patterns of expression and coinducibility during iron overlaod[J]. FEBS Lett,2003,542(1-3):22-26.
Ishida T, Zheng Z, Dichek Ice, et al. Molecular cloning of nonsecreted endothelial cell-derived lipase isofonns[J]. Genomics.2004 Jan,83(1):24～33
Jain C. Gene,1993; 133(1):99-102.
Jia HP, Schuae BC, Schudy A, et al. Discovery of new human β-defensins using a genomics base dapproach[J]. Gene,2001,263(122):211-218.
Jin FL, et al. Expression and characterization of antimicrobial peptide CecropinAD in the methylotrophic yeast Pichia pastoris[J]. Process Biochem (2008),doi:10.1016/j.procbio.2008.08.012
Jolles P, Jolles J. What's new in lysozyme research? Always a model system, today as yesterday[J]. Mol Cell Biochem 1984;63:165-89.
Kaattari SL, Piganelli JD. The specific immune response:humoral defense[M]//Iwama G, Nakanishi T. The fish immune system,organism,pathogen and environment.San Diego USA:Academic Press,1996:207～254.
Kachamakova NM, Irnazarow I, Parmentier HK, et al. Genetic differences in natural antibody levels in common carp (Cyprinus carpio L.) [J]. Fish and Shellfish Immunology 2006;21:404～13.
Katsumi Matsuzaki. Why and how are peptide-lipid interactions utilized for self-defense? Magainins and tachyplesins as archetypes 。 Biochimica et Biophysica Acta (BBA)-Biomembranes, Volume 1462, Issues 1-2,15 December 1999, Pages 1～10
Kimbrell D A, Bender B. The evolution and genetics of innate immunity[J].Natuer Rev.Genet.2001;2:256～267
Kimura Y., Madhavan M., Call MK., et al. Expression of complement 3 and complement 5 in newt limb and lens regeneration[J]. Journal of Immunology 2003;170:2331-9.
Kos J., Werle B., Lah T., Brunner N. Cysteine proteinases and their inhibitors in extracellular fluids:markers for diagnosis and prognosis in cancer[J]. Int J Biol Markers,2000; 15:84-89.
Kothary R.K., Jones. D., Candido E.P.70-kilodalton heat shock poly-peptides from rainbow trout:characterization of cDNA sequences[J]. Mol Cell Biol 1984;4:1785-1791.
Krause A, Neitz S, Magert HJ, et al. LEAP21, a novel highly disulfide bonded human peptide,exhibits antimicrobial activity[J]. FEBS Lett,2000;480:147～ 150.
Kulaksiz H, Theilig F, Bachmann S, et al. The iron-regulatory Peptide hormone hepcidin:expression and cellular localization in mammalian kidney[J] Journal of Endocrinolog,2005,184:361～370.
Lange S, Bambir S, Dodds AW, Magnado'ttir B. The ontogeny of complement component C3 in Atlantic cod (Gadus morhua L.)-an immunohistochemical study[J]. Fish and Shellfish Immunology,2004; 16:359～67.
Langston AL, Johnstone R, Ellis AE. The kinetics of the hypoferraemic response and changes in levels of alternative complement activity in diploid and triploid Atlantic salmon, following injection of lipopolysaccharide[J]. Fish and Shellfish Immunology,2001; 11:333～45.
Langston AL., Bricknell IR., Ellis AE. Iron binding capacity of peripheral blood leucocyte lysates from Atlantic salmon (Salmo salar L.). In:Barnes AC, Davidson GA, Hiney MP, McIntosh D, editors. Methodology in fish diseases research. Aberdeen:Fisheries Research Services,1998; p.111～6.
Lele Z, Engel S, Krone P.H. Hsp47 and hsp70 gene expression is differentially regulated in a stress and tissue-specific manner in zebrafish embryos[J]. Dev Genet 1997;21:123-133.
Lemaitre B.The road to Toll[J]. Nat Rev Immunol.2004;4(7):521～527.
Lie O, Evensen O, Sorensen A, Froysadal E. Study on lysozyme activity in some fish species[J]. Dis Aquat Org 1989;6:1～5.
Lindsay GJH., Gooday GW. Chitinolytic enzymes and the bacterial microflora in the digestive tract of cod, Gadus morhua[J]. J Fish Biol 1985;26:255～65.
Llanos RJ, Whitacre CM, Miceli DC. Potential involvement of C3 complement factor in amphibian fertilization[J]. Comparative Biochemistry and Physiology, Part A 2000;127:29～38.
Lomax J, McCray AT. Mapping the Gene Ontology into the Unified Medical Language System[J].Comp Funct Genomics,2004;5:345～361.
Lovoll M, Kilvik T, Boshra H, et al. Maternal transfer of complement components C3-1, C3-3, C3-4, C4, C5, C7, Bf, and Df to offspring in rainbow trout (Oncorhynchus mykiss) [J]. Immunogenetics,2006;58:168～79.
Magnadottir B, Gudmundsdottir S, Gudmundsdottir BK. Study of the humoral response of Atlantic salmon (Salmo salar L.), naturally infected with Aeromonas salmonicida ssp achromogenes[J]. Veterinary Immunology and Immunopathology,1995; 49:127～42.
Magnadottir B, Lange S, Steinarsson A, Gudmundsdottir S. The ontogenic development of innate immune parameters of cod (Gadus morhua L.) [J]. Comp Biochem Physiol B Biochem Mol Biol,2004; 139:217～24.
Manson FDC., Fletcher TC., Gooday GW. Localization of chitinolytic enzymes in blood of turbot, Scophthalmus maximus, and their possible roles in defense[J]. J Fish Biol,1992;40:919～27.
McGinnis S, Madden TL.BLAST:at the core of a powerful and diverse set of sequence analysis tools.Nucleic Acids Research,2004;32:W20-W25.
Melchedov S, de Ilarduya OM,Ohlrogge J.Towards a functional catalog of the plant genome:a survey of gene for lipid biosynthesis[J]. Plant Physiol, 2000;122(2):389-402.
Molina A, Biemar F, Muller F, et al.Cloning and expression analysis of an inducible HSP70 gene from tilapia fish[J].FEBS Lett,2000;474:5-10.
Moon JY, Henzler-Wildman KA, Ramamoorthy A. Expression and purification of a recombinant LL-37 from Escherichia coli[J]. Biochim Biophys Acta,2006;1758(9):1351-1358.
Morgan BP, Marchbank KJ, Longhi MP, et al. Complement:central to innate immunity and bridging to adaptive responses[J]. Immunological Letters, 2005;97:171-9.
Murphy JE, Robert C, Kupper TS. Interleukin-land cutaneous inflammation:a crucial link between innate and acquired immunity [J]. J Invest Dermatol,2000;114(3):602-608.
Mwthy TV. Expression of GST-fused kinase domain of human Csk homologous kinase from pichia pastoris facilitates easy purification[J]. Biotechnol Let,2004; 26(5):443-449.
Nam BH, Yamamoto E,Hirono I, et al. A survey of expressed genes in the leukocytes of Japanese founder, Paralichthy solivaceus, infected with Hiramerhabdovirus[J] Dev Comp Immunol,2000;24(1):13
Nemat 0 K, Saul R. The chitin catabolic cascade in the marine bacterium Vibrio fumissii[J]. JBiol Chem,1996;271(52):33425-33432.
Nemeth E, Rivera S, Gabayan V, et al. IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin[J]. J Clin Invest,2004; 113:1271.
Nicolas G, Chanvet C, Viatte L, et al. The gene encoding the iron regulatory peptide hepcidin is regulated by anemia, hypoxia, and inflammation[J]. The Journal of Clinical Investigation,2002,110,1037～1044.
Nicolas G, Bennoun M, Devaux I, et al. Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2)knockout mice[J]. Proc Natl Sci USA,2001;98:8780～8785.
Nicolas G, Bennoun M, Porteu A, et al. Severe iron deficiency amernia in transgenic mice expressing liver hepcidin[J]. Proceedings of the National Academy of Sciences of the United States of America,2002,99:4596-4601.
Noga E J,Fan Z, Silphaduang U. Histone-like proteins from fish are lethal to the parasitio dinoflagellate Amyloodinium ocellatum[J]. Parasitology,2001; 123(Ptl):57-65.
Nollen E.A.A., Morimoto R.I. Chaperoning signaling pathways:molecular chaperones as stress-sensing "heat shock" proteins. J Cell Sci 2002; 115:2809-2816. Takao S., Tetsuo K, Katsumasa G., et al.Possible role of calcineurin in heating-related increase of rat muscle mass. Biochemical and Biophysical Research Communications,2005;331(4):1301-1309.
Nonaka M, Smith SL. Complement system of bony and cartilaginous fish[J]. Fish and Shellfish Immunology,2000;10:215-28.
Oren Z, Shai Y. A class of highly potent antibacterial peptides derived from pardaxin:a pore-forming peptide isolated from moses sole fish Pardachirus marmoratus[J].r J Biochem,1996;237(1):303-310.
Owhashi M., Arita H., Hayai N. Identification of a novel eosinophil chemotactic cytokine (ECF-L) as a chitinase family protein[J]. J Biol Chem,2000; 275(2):1279-86.
P.C.特纳,A.G.迈克伦南,A.D.贝茨,M.R.H.怀特,等著；刘进元,李文君,王薛林等译校.分子生物学(第二版)[M]．北京：科学出版社,2004.
Parham P., Ohta T. Population biology of antigen presentation by MHC class I molecules[J]. Science,1996;272:67～74.
Park CH, Valore EV, Waring AJ, et al. Hepcidin, a urinary antimicrobial peptide synthesized in the liver[J]. Biol Chem,2001;276:7806-7810.
Park IY, Park CB, Kim MS, et al. Parasin I, an antimicrobial peptide derived from histone H2A in the catfish, Parasilurus asotus[J]. FEBS Lett,1998;437 (3):258～262.
Park K C, Osborne J A, Tsoi S C M, et al. Expressed sequence tags analysis of Atlantic halibut Hip poglossuship poglossus liver, kidney and spleen tissues following vaccination against Vibrio anguil larum and A eromonas salmonicida[J]. Fish Shellfish Immunol,2005;18:393～415.
Paul Y, Kassebaum C, Lazarovici P,et al. Translocation of acylated pardaxin into cells[J]. FFBS Letters,1998;440(122):131-134.
Peng L, Xu Z, Fang X, et al. High—level expression of soluble human β-defensin-2 in Escherichia coli[J]. Process Biochem,2004,39(12):2199.
Persson A.C., Stet.R.JM., Pilstrom L., et al. Characterization of MHC class 1 and beta2-microglobulin sequences in Atlantic cod revealing an unusual high number of expressed class I genes[J].Immunogenetics,1999;50(1):49～59.
Pigeon C, Ilyin G, Courselaud B, et al. A new mouse liver-specific gene, encoding a protein homologous to human antimicrobial peptide hepcidin, is overexpressed during iron overload[S].J Biol Chem,2001,276(11):7811～ 7819.
Pouny Y, Rapaport D, et al. Interaction of antimicrobial dermaseptin and its fluorescently labeled analogues with phospholipid membranes [J]. Biochemistry,1992,31(49):12416-12423.
Press MCL, Dannevig BH, Landsverk T. Immune and enzyme histochemical phenotypes of lymphoid and nonlymphoid cells within the spleen and head kidney of Atlantic salmon (Salmo salar L.) [J]. Fish and Shellfish Immunology,1994;4:79-93.
Qiu Z, Hingley ST, Simmons G, et al. Endosomal proteolysis by cathepsins is necessary for murine coronavirus mouse hepatitis virus type 2 spike-mediated entry[J]. J Virol,2006;80:5768-76.
Raldu'a D, Fabra M, Bozzo MG, et al. Cathepsin B-mediated yolk protein degradation during killifish oocyte maturation is blocked by an HATPase inhibitor:effects on the hydration mechanism[J]. Am J Physiol Regul Integr Comp Physiol,2006;290:456～466.
Rao XC, Li S, Hu JC, et al. A novel carrier molecule for high-level expression of peptide antibiotics in Escherichia coli[J]. Pro Exp Purif,2004,36:11～18.
Robertsen B. The interferon system of teleost fish[J]. Fish and Shellfish Immunology,2006;20(2):172-191.
Roeder T. Solid-phase cDNA library construction, a versatile apporach[J]. Nucleic Acids Res,1998,26(14):3451-3452.
Russell S., Lumsden JS. Function and heterogeneity of fish lectins[J]. Veterinary Immunology and Immunopathology,2005; 108:111～120.
Salte R, Gjoen HM, Norberg K, et al. Plasma protein levels as potential marker traits for resistance to furunculosis[J]. J Fish Dis,1993;16:561～8.
Sebastian P, Wallwits J, Schmidt S. Semi automated production of a set of different recombinant GST-Streptag fusion protein[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2003,786(1-2):343～355.
Shai Y. Mechanism of the binding, insertion and destabilization of phospholipid bilayer membranes by a-helical antimicrobial and cell non-selective membrane-lytic peptides[J].Biochim Biophys Acta,1999;1462:55～70.
Shai Y. Pardaxin:channel formation by a shark repellant peptide from fish[J]. Toxicology,1994,87(13):109～129.
Sharmeks A D. Gene,1994; 138(1-2):105-108.
Sheridan M A. Lipid dynamics in fish:aspects of absorption, transportation, deposition and mobilization[J]. Comp. Biochem. Physiol,1988,90(4): 670～690.
Sherwin T, Brookes NH, Loh IP, Poole CA, Clover GM. Cellular incursion into Bowman's membrane in the peripheral cone of the keratoconic cornea[J]. Exp Eye Res 2002;74:473～482.
Shike H, Lauth X, Westerman M E, et al. Bass hepcidin is a novel antimicrobial peptide induced by bacterial challenge[J]. Eur J Biochem,2002,269(8): 2232～2237.
Shike H, Shimizu C, Lauth X, et al. Organization and expression analysis of the zebrafish hepcidin gene, antimicrobial peptide gene conserved among vertebrates[J]. Dev Comp Immunol,2004,28:747～754.
Shona K, Whyte. The innate immune response of finfish-A review of current knowledge[J]. Fish and Shellfish Immunology,2007;23:1127-1151.
Silphaduang U, Edwardb JN. Peptide antibiotics in mast cells of fish[J].Nature,2001,414(6861):268-269.
Sommer S. The importance of immune gene variability (MHC) in evolutionary ecology and conservation[J]. Front. Zool.2005;2:1～18.
Song LC, Xiang SJ, Guo CY, et al. Cloning, characterization, and expression analysis of hepcidin gene from Red Sea bream(Chrysophrys major)[J]. AAC, 2005,49(4):1608-1612.
Stafford JL, Belosevic M. Transferrin and the innate immune response of fish: identification of a novel mechanism of macrophage activation[J]. Dev Comp Immunol,2003;27:539-554.
Stafford JL, Neumann NF, Belosevic M. Products of proteolytic cleavage of transferrin induce nitric oxide response of goldfish macrophages[J]. Dev Comp Immunol,2001;25:101-15.
Sugano S, Maruyama K. Toward the construction of a full length cDNA library[J]. Tanpakushitsu Kakusan Koso,1993,38:476～484.
Sultmann H., Mayer W.E., Mayer F. Organization of MHC class II B genes in the zebra fish(Brachydanio reio) [J].Genomics 1994;23(1):1～4.
Susan E D, et al. Identification and expression analysis of hepcidin-like antimicrobial peptide in bony fish [J]. Developmental and Comparative Immunology,2003,27:589～601.
Suzuki M,Morimatsu M, Yamashita T,et al. A novel serum chitinase that is expressed in bovine liver[J]. FEBS Lett.2001;506(2):127-130.
Takeuchi H, Figueroa F, O'hUigin C, et al.Cloning and characterization of class I Mhc genes of the zebrafish, Danio rerio[J]. Immunogenetics,1995;42(2):77～ 84.
Tateno H, Saneyoshi A, Ogawa T, et al. Isolation and characterization of rhamnose-binding lectins from eggs of steelhead trout (Oncorhynchus mykiss) homologous to low density lipoprotein receptor superfamily[J]. Journal of Biological Chemistry 1998;273(30):19190-7.
Tort L, Balasch JC, Mackenzie S. Fish immune system. A crossroads between innate and adaptive responses[J]. Immunologi'a 2003;22(3):277-86.
Townes CL, Michailidis G, Nile CJ, et al. Induction of cationic chicken liver-expressed antimicrobial peptide 2 in response to Salmonella enterica infection[J]. Infect Immun,2004,72(12):6987～6993
Townsend, A., and H. Bodmer. Antigen recognition by class I-restricted T lymphocytes[J]. Annu. Rev. Immunol,1989;7:601.
Vanerp S H M., Dixon B, Figneroa F, et al. Identification and characterization of new major histocompatibility complex class I gene in carp (Cyrinus carpio) [J]. Immunogenetics,1996a;44(1):49-61.
Vanerp S.H.M., Egberts E., Stet R.J. Characterization of major histocompatibility complex class ⅡA and B genes in gynogenetic carp clone[J]. Immunogenetics, 1996b,44(3):192～202.
Vilain C, Wetzel M-C, Du Pasuier L, et al. Structural and functional analysis of spontaneous anti-nitrophenyl antibodies in three cyprinid fish species:carp (Cyprinus carpio), goldfish (Carassius auratus) and tench (Tinca tinca) [J]. Developmental and Comparative Immunology,1984;8:611-22.
Wang RF, Wang X, Johnston SL, et al. Development of a retrovirus-based complementary DNA expression system for the cloning of tumor antigens[J].Cancer Res,1998,58 (16):3519-3525.
Wang Z, Zhang S, Wang G, et al. Complement activity in the egg cytosol of zebrafish Danio rerio:evidence for the defense role of maternal complement components[J]. Plos One,2008;3(1):1463
Weinstein D A, Roy C N, Fleming M D,et al. Inappropriate expression of hepcidin is associated with iron refractory anemia:Implications for the anemia of chronic disease[J]. Blood,2002,100:37-76.
Yang M, Wang KJ, Zhou HL. A novel hepcidin-like antimicrobial peptide isolated from the Japanese Bass[J]. J Xiamen University (Natural Science),2004, 43(3):286～287.
Yano T. The nonspecific immune system:humoral defense. In:Iwama G, Nakanishi T, editors. The fish immune system:organism, pathogen, and environment. San Diego, USA:Academic Press;1996,p.105-53.
Yeaman MR, Yount NY. Mechanisms of antimicrobial peptide action and resistanc[J]. Pharmacol.Rev.2003;55 (1):27～55
Yifeng Li, Xia Li, Guangshun Wang.Cloning, expression, isotope labeling, and purification of human antimicrobial peptide LL-37 in Escherichia coli for NMR studies[J]. Protein Expression and Purification,2006,47:498-505.
Yousif AN., Albright LJ., Evelyn TPT. In vitro evidence for the antibacterial role of lysozyme in salmonid eggs[J]. Dis Aquat Org,1994;19:15-9.
Zasloff. Antimicrobial peptides of multicellular organisms[J]. Nature, 2002;415:389-395.
Zhang L, Falls T, Wu M, et al.Determinants of recombinant productionof antimicrobial cationic peptides and creation of peptide variant inbacteria[J].Biochem Biophys. Res. Commun.1998,247:674～680.
Zhang Y A, Zou J, Chang C I, et al. Discovery and characterization of two types of liver-expressed antimicrobial peptide 2(LEAP-2)genes in rainbow trout[J]. Vet Immunol Immunopathol,2004,101:259～269.
Zhao, D X, Song S H, Wang Q, et al., Discovery of immune-related genes in Chinese mitten crab (Eriocheir sinensis) by expressed sequence tag analysis of haemocytes[J]. Aquaculture,2009,283 (3-4):297-303.
Zugel U, Kaufmann S.H.E. Role of Heat Shock Proteins in Protection from and Patho genesis of Infectious Diseases. Clin. Microbiol. Rev,1999; 12:19～ 39.
Zuo X, Woo PTK. In vivo neutralization of Cryptobia salmositica metallo-protease by alpha 2-macroglobulin in the blood of rainbow trout Oncorhynchus mykiss and in brook charr Salvelinus fontinalis[J]. Dis Aquat Org, 1997;29:67-72.
陈文波,刘黎黎,李文笙,等.鲤鱼肝胰脏cDNA文库的表达序列标签分析及Lb-Fabp mRNA的特征[J].水生生物学报,2009,33(3)：376～384。
方永强,中庆祥,翁幼竹,等.文昌鱼头部cDNA文库的构建[J].台湾海峡,2003,22(2)：131-135.
胡国良,曹华斌,郭小权,等.高能低蛋白日粮对蛋鸡肝载脂蛋白A-I基因mRNA表达的影响[J].中国兽医科学2009,39(7)：641-645.
胡松年主编.基因表达序列标签(EST)数据分析手册[M].杭州：浙江大学出版社,2005.
姜秀云.牛分枝杆菌主要保护性抗原基因的克隆、表达及免疫研究[D].吉林农业大学博士论文,2005.
赖玉平,彭沂非,郁正艳,等.DCD-1L在毕赤酵母中的克隆和表达[J].中国生物工程杂志.2004,24(2)：61-65.
李伟,陈松林.牙鲆抗菌肽hepcidin基因在大肠杆菌中的融合表达[J].高技术通讯.2007,17(7)：765-770.
李旭霞,逢越,肖蓉,等.日本七鳃鳗(Lampetrs japonica)唾液腺cDNA文库的构建[J].辽宁师范大学学报(自然科学版),2004,27(1)：73～75.
李志琼,余冰,张克英,等.饲粮中添加a-亚麻酸对产蛋鸡载脂蛋白B和血脂的影响[J].动物营养学报,2008,20(6)：674-678.
刘家芳,杨代勤,阮国良.黄鳝消化酶活性的研究[J].湖北农业科学,2007,46(5)：804-807.
彭彩云.黄鳝养殖常见病害的防治[J].现代农业科技,2009,14：304～305
申艳敏,魏建超,尚书文等.人源抗菌肽LL-37在毕赤酵母中的高效表达及其活性检测[J].微生物学通报.2008,35(4)：539～544.
唐勇.黄鳝抗菌活性物质的制备及其抑菌效果的研究[D].四川农业大学硕士论文,2007.
王克坚,周红玲,杨明.海水养殖鲈鱼分离出一种hepcidin抗菌肽新基因[J].厦门大学学报：自然科学版,2004,43(3)：286～287.
吴乃虎.基因工程原理(第二版)[M].北京：科学出版社,1999.
徐奇友,单安山,王安.乳铁蛋白对早期断奶仔猪组织中微量元素含量的影响 [J].动物营养学报,2005,17(4)：62.
杨芳,贺智敏,詹显全,等.大鲵肝脏组织定向cDNA文库的构建及鉴定[J].动物学报,2004,50(3)：475-478.
杨贵贞等著.免疫活性细胞的效应[M].北京：科学出版社,1986
杨明,王克坚,周红玲,等.我国海水养殖真鲷分离出hepcidin抗菌肽新基因[J].高技术通讯,2004,14(增刊)：323～328.
杨旭.抗菌肽Pexiganan和IB-367基因在大肠杆菌中的克隆与表达[D].天津大学硕士论文.2006
药立波,常智杰,马文丽.医学分子生物学实验技术[J].北京：人民卫生出版社,2002.164.
尹娜,李鸿钧,彭梅等.抗菌肽Cecropin D在毕赤酵母中的表达、纯化及活性鉴定[J].中国生物制品学杂志,2008,21(3)：185～189.
张勇,张为民,李欣,等.石斑鱼卵巢cDNA文库构建及脂肪酸结合蛋白克隆[J].中山大学学报(自然科学版),2003,42(2)：66-69.
张宁.大海马cDNA文库构建、分析及抗菌肽新基因的初步功能研究[D],中山大学博士论文,2007.
张学俊,屈刚,朱文漓,等.草鱼肠道cDNA文库构建及部分ESTs分析[J].水生生物学报,2007,31(2)：251-258.
张雅丽,杨国庆,郭英,陈永福.密码子优化的牛精蛋白基因在大肠杆菌中的表达[J].高技术通讯,2002，4(1)：42～46.