赤点石斑鱼(<I>Epinephelus akaara</I>)MHC ⅡB基因的克隆与表达多态性分析

丁少雄; 张之文; 杜佳莹; 王军; 曾华嵩; 王颖; 陈晓峰

赤点石斑鱼(Epinephelus akaara)MHC ⅡB基因的克隆与表达多态性分析

1.
厦门大学, 海洋与环境学院, 福建, 厦门, 361005
2.
国家海洋局, 第三海洋研究所, 福建, 厦门, 361005

基金项目: 福建省自然科学基金(2060203);福建省新世纪人才资助计划

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出版历程
- 收稿日期: 2008-08-07
- 修回日期: 2009-01-21

The cloning and polymorphism of MHC class ⅡB gene from Epinephelus akaara

1.
Department of Oceanography and Institute of Subtropical Oceanography, Xiamen University, Xiamen 361005, China
2.
Third Institute of Oceanoqraphy, State Oceanic Administration, Xiamen 361005, China

摘要

摘要: 主要组织相容性复合体(Major histocompatibility complex,MHC)是脊椎动物体内重要的非特异性免疫基因之一。为了研究赤点石斑鱼的MHC基因的结构与表达特性,本研究运用RACE,Realtime PCR及SSCP等技术,首次成功克隆获得赤点石斑鱼(Epinephelus akaara)非特异性免疫因子MHC ⅡB基因的4个cDNA全序列,序列全长为1 195~1 355 bp,含有3'UTR、启动子、多肽结合区(β1)I、GC区(β2)、跨膜区、胞质区和5'UTR区,编码区大小为750 bp。氨基酸序列分析发现赤点石斑鱼MHC ⅡB分子具有经典MHC蛋白分子的空间结构,在多肽结合区存在极其丰富的变异,β1区由1个α螺旋与4个反向平行的β折叠构成多肽结合区,β2区由多个β折叠构成2个反向平行的三明治结构。利用SSCP技术研究了赤点石斑鱼MHC ⅡB的表达多态性,发现其在头肾、心、肝等12个组织器官中均能有效表达,表达多态性也异常丰富。此外,根据MHCIIB分子中相对保守的IGC区氨基酸序列构建了脊椎动物的系统进化树,也表明了该分子的IGC区氨基酸序列可以作为研究鱼类物种间进化关系的良好标记。
- 赤点石斑鱼 /
- MHCⅡB基因 /
- 多态性
Abstract: Major histocompatibility complex is one of crucial innate immune factors in vertebrates.In this study, RACE, Realtime PCR and SSCP techniques had been used to study the structure and expression characterization o f MHC gene from Epinephelus akaara, 4 complete cDNA sequences of MHC class Ⅱβ chain from E.akaara was obtained, the sequences have the length of 1195~1355 bp, and contain a 3' UTR, promoters, a peptide-binding region(β1), an immuno globulin-like region(β2), a transmembrane region, a cytoplas ic region, and 5' UTR.The size of their coding region is 750 bp, exhibits a classic MHC 3D molecular structure and abundant polymorphisms in the peptide-binding region.Within the peptide-binding region,β1 included an alpha-helical region and a beta sheet of four strands in antiparallel orientation, while β2 forms a sandwhich-like structure made of two ant iparallel sheets, with each containing several strands.Using SSCP technique, we observed high levels of expression of poly morphism in 12 tissue such as head-kidney, heart and liver.We also constructed phylogenetic tree based on Neighbour-Joining methods with IGC region of the MHC ⅡB gene.These results showed that the amino acid sequence of the IGC region of MHC ⅡB is informative for phylogenetic studies in fishes.
- Epinephelus akaara /
- MHC class Ⅱ gene /
- polymorphism

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参考文献(31)

王重庆.分子免疫学基础[M].北京:北京大学出版社,1997:139-140.

PAUL W E Fundsmental Immunology.Fourth Edition[M].Lippincott-Raven Publishers,Philadelphia,1999:297-298.

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.

EDWARD S V,HEDRICK P W.Evolution and ecology of MHC molecules:from genomics to sexual selection[J].Trends in Ecology and Evolution,1998,13:305-311.

HEDRICK P W,KIM T J.Genetics of complex polymorphisms:parasites and maintenance of MHC variation[M]//SINGH R S,KRIMBAS C K.Evolutionary Genetics From Molecules to Morphology.New York,Cambridge University Press,1999.

陈信忠,苏亚玲,龚艳清.闽南地区养殖石斑鱼病毒性神经坏死病初步研究[J].福建水产,2003,9:11-14.

吴后波,吴灶和,潘金培.海水网箱养殖赤点石斑鱼溃疡病致病菌研究[D]//热带海洋研究(五).北京:科学出版杜,1997:91-95.

杨霞,吴信忠.赤点石斑鱼的普通变形菌病原学研究[J].水产科学,2005,(24):5-7.

BINGULAC P J,FIGUEROA F.SATO A,et al.Mapping of MHC class Ⅰ and class Ⅱ regions to different linkage groups in the zebrafish.Daniorerio[J].Immunogenetics,1997,46:129-144.

JUUL-MADSEN H R,GLUMANN J,MADSN H O,et al.MHC class Ⅱ beta-chain expression in the rainbow trout[J].Seen J of Immunol,1992,35:687-694.

KOPPANG E O,LUNDIN M,PRESS C M,et al.Differing levels of Mhc class Ⅱβ chain expression in a range of tissues from vaccinated and non-vaccinated Atlantic salmon(Salmo salar L.)[J].Fish & Shellfish Immunology,1998,8:183-196.

KOPPANG E O,PRESS C M,RONNlNGEN K,et al.Expression of Mhc class Ⅰ mRNA in tissues from vacdnated and nonvaccinated Atlantic salmon(Salmo udar L.)[J].Fish & Shellfish Immunology,1998,8:577-587.

HEEMSTRA P C,RANDALL J E.Groupers of the World[M].Rome:FAO Species Catalogue,1993.

成庆泰,郑葆珊.中国鱼类系统检索(上册)[M].北京:科学出版社,1987.

孟庆闻,苏锦祥,缪学祖.鱼类分类学[M].北京:中国农业出版社,1995.

SAMBROOK J,FRITSCH E F,MANIATIS T.Molecular Cloning:A Laboratory Manual[M].Now York:Cold Spring Harbor Laboratory Press,1989.

COSSON P,BONIFACINO J S.Role of transmembrane domain interactions in the assembly of class Ⅱ MHC molecules[J].Science,1992,258:659-662.

RODRIGUES P N S.Expression of Major histocompatibility complex genes in carp(Cyprinus carpio L.)[D].Dutch:Elsevier Science Ltd and Academic Press Ltd,1996:1-27.

周光炎.免疫学原理[M].上海:上海科学技术出版社,2000,63-121.

RENÉ J M.Inference of structure and function of fish major histocompatibility complex(MHC),molecules from expressed genes[J].Fish & Shellfish Immunology,1996,6:305-318.

金伯泉.细胞和分子免疫学[M].西安:世界图书出版公司,1998:213-258.

ONO H,O'HUIGIN C,VINCEK V,et al.New β chain-encoding MHC classⅡgenes in the carp[J].Immunogenetics,1993,38:146-149.

RODRIGUES P N S,HERMSEN T T,TOMBOUT J H,et al.Detection of MHC classⅡtranscripts in lymphoid tissues of the common carp(Cyprinus carpio L.)[J].Dev Comp lmmunol,1995,19:483-495.

张玉喜,陈松林.大菱鲆MHCⅡB基因全长cDNA的克隆与组织表达分析[J].高技术通讯,2006,16(8):859-863.

VLADIMIR V,DAGMAR K,GRASER R T.et al.Molecular cloning of major histocompatibility complex class Ⅱ B gene cDNA from the Bengalese finch Lonchura striata[J].Immunogenetics,1995,42(4):262-267.

O'RIEN S J.Comparative genome organization of the major histocompatibility complex:lessons from the Felidae[J].Immunological Reviews,1994,167:133-144.

KRISTEN D A,GIESE A R,HOLLY L M,et al.Resistance to three pathogens in the endangered winter-run chinook salmon(Oncorhynchus tshawytscha):effects of inbreeding and major histocompatibility complex genotypes[J].Can J Fish Aquat Sci,2002,59(6):966-975.

KRZYSZTOF L R,GEERT F W,MIKOLAJ A,et al.Application of PCR-RF-SSCP to study major histocompatibility class Ⅱ Bpolymorphism in common carp(Cyprinus carpio L.)[J].Fish & Shellfish Immunology,2008,24:734-744.

SATO A,FLIGUEROA F,MURRAY B W,et al.Nonlinkage of major histocompatibility complex class Ⅰ and classⅡloci in bony fishes[J].Immunogenetics,2000,51:108-116.

YUKO O,KAZUHIKO O,MCKINNEY E.Primitive synteny of vertebrate major histocompatibility complex class Ⅰand classⅡgenes[J].PNAS,2000,97:4712-4717.

SIMONA B,DAVID B,IRVING L W.Molecular Evolution of the Vertebrate Immune System[J].PNAS,1994,91(23):10769-10770.

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