韩进刚,付小哲,石存斌,李凯彬,潘厚军,吴淑勤

• 1:中国水产科学研究院珠江水产研究所
• 2:中国水产科学研究院珠江水产研究所
• 3:中国水产科学研究院珠江水产研究所
• 4:中国水产科学研究院珠江水产研究所

摘要(Abstract)：

以免疫两周的剑尾鱼为材料,提取脾脏总RNA,通过逆转录-聚合酶链式反应(RT-PCR)扩增出分泌型免疫球蛋白M(secretory immunoglobulin M,sIgM)重链基因的核心序列,再应用3’和5’快速扩增cDNA末端(RACE)方法扩增其末端序列,拼接后获得剑尾鱼sIgM重链全长cDNA序列。全长序列含有免疫球蛋白的信号序列:FKCIANH,得到的5个可变区序列可能来源于4个不同的VH家族。经用半定量RT-PCR分析sIgM在脾脏和头肾的表达变化,初步认为利用剑尾鱼头肾sIgM的mRNA表达变化替代检测血清抗体效价具有可能性。

关键词(KeyWords)： 剑尾鱼;免疫球蛋白M;基因表达;基因克隆

Abstract:

Keywords:

基金项目(Foundation): 科技部社会公益项目(2004DIB1J029);; 国家科技支撑计划项目(2006BAD03B05)

作者(Author): 韩进刚,付小哲,石存斌,李凯彬,潘厚军,吴淑勤

参考文献(References)：

• [1]吴淑勤,黄志斌,石存斌,等.水生实验动物——剑尾鱼[M].北京:中国农业出版社,2005:7-9;93-108.
• [2]雄清明,陆承平.迟缓爱德华菌(Edwardasiella tarda)对小鼠和剑尾鱼的免疫保护试验[J].中国兽医学报,2002,22(3):251-253.
• [3]张吉红,储卫华,陆承平.口服嗜水气单胞菌生物被膜疫苗的动物免疫试验[J].中国水产科学,2004,11(5):404～407.
• [4]Nakao M,Moritomo T,Tomana M,et al.Isolation of cDNA encoding the constant region of the immunoglobulin heavy-chain from common carp(Cyprinus carpio L.)[J].Fish&shellfish immunology,1998,8(6):425–434.
• [5]Kabat E A,Wu T T,Perry H M,et al.Sequences of proteins of immunological interest[M].Washington:US Department of health and human services,1991:320-790.
• [6]Lundqvist M L,Strmberg S,Pilstr?m L.Ig heavy chain of the sturgeon Acipenser baeri:cDNA sequence and diversity[J].Immunogenetics,1998,48(6):372-382.
• [7]Nil Ratan Saha,Hiroaki Suetake,Yuzuru Suzuki.Analysis and characterization of the expression of the secretory and membrane forms of IgM heavy chains in the puffer fish,Takifugu rubripes[J].Molecular immunology,2005,42(1):113-124.
• [8]Hansen H,Leong J A,Kaattari S.Complete nucleotide sequence of a rainbow trout cDNA encoding a membrane-bound form of immunoglobulin heavy chain[J].Mol Immunol,1994(31):499-501.
• [9]Bengten E,Leanderson T,Pilstrom L.Immunoglobulin heavy chain cDNA from the teleost Atlantic cod(Gadus morhua L.):nucleotide sequences of secretory and membrane form show an unusual splicing pattern[J].Eur J Immunol,1991,21(12):3027-3033.
• [10]Nakao M,Moritomo T,Tomana M,et al.Isolation of cDNA encoding the constant region of the immunoglobulin heavy-chain from common carp(Cyprinus carpio L.)[J].Fish&shellfish immunology,1998(8):425-434.
• [11]Espelid S,Halse M,Solem S T,et al.Immunoglobulin genes and antibody responses in the spotted wolfish(Anarhichas minor Olafsen)[J].Fish&shellfish Immunol,2001,11(5):399-413.
• [12]Kokubu F,Hinds K,Litman R,et al.Complete structure and organization of immunoglobulin heavy chain constant region genes in a phylogenetically primitive vertebrate[J].The EMBO journal,1988,7(7):1979-1988.
• [13]Yang F,Ventura-Holman T,Waldbieser G C,et al.Structure,genomic organization,and phylogenetic implications of six new VH families in the channel catfish[J].Mol Immunol,2003,40(5):247-260.
• [14]Frazer K L,Capra J D.Immunoglobulins:structure and function[M]//Paul WE,ed.Fundamental Immunology.4th ed.New York:Lippincott-Raven press,1999:37-74.
• [15]Amemiya C T,Litman G W.Complete nucleotide sequence of an immunoglobulin heavy-chain gene and analysis of immunoglobulin gene organization in a primitive teleost species[J].Proc Natl Acad Sci USA,1990,87(2):811-815.
• [16]王荻,刘红柏.5种鲟鱼免疫球蛋白重链恒定区序列研究[J].遗传,2006,28(10):1247-1253.
• [17]Cheng C A,John J A,Wu M S,et al.Characterization of serum immunoglobulin M of grouper and cDNA cloning of its heavy chain[J].Veterinary immunology and immunopathology,2006,109(3):255-265.
• [18]Shakin-Eshleman S H,Spitalnik S L,Kasturi L.The amino acid at the X position of an Asn–X–Ser sequon is an important determinant of N-linked core-glycosylation efficiency[J].J Biol Chem,1996,271(11):6363–6366.
• [19]de Lalla C,Fagioli C,Cessi F S,et al.Biogenesis and function of IgM:the role of the conserved mchain tailpiece glycans[J].Mol Immunol,1998,35(13):837-845.
• [20]李亚南,王冀平,邵健忠,等.鱼类免疫应答中的温度效应研究[J].浙江大学学报(理学版),1999(3):87-91.