凡纳滨对虾肌肉组织cDNA全长文库的构建  

Construction of the full length cDNA library from muscular tissue of Litopenaeus vannamei

作　　者：熊建华[1] 高永华[1,2] 马宁[1] 盛小伟[1,2] 陈晓汉[1] 

机构地区：[1]广西水产研究所遗传育种与健康养殖重点实验室,南宁530021 [2]桂林理工大学化学与生物工程学院,广西桂林541000

出　　处：《南方农业学报》2011年第1期86-89,共4页Journal of Southern Agriculture

基　　金：国家科技支撑计划项目(2008BADB9B03);国家虾产业技术体系岗位科学家经费项目(nycytx-46);广西直属公益性科研院所基本科研项目(2060302GXIF-2008-02);广西自然科学基金项目(桂科自0991106)

摘　　要：【目的】为了在短期内获得大量凡纳滨对虾肌肉组织的功能基因表达信息,为深入了解功能基因在凡纳滨对虾肌肉组织中的表达提供分子生物学依据。【方法】通过构建凡纳滨对虾肌肉组织的全长cDNA文库,并进行EST测序分析。【结果】文库质量分析表明,初始文库库容约8.50×106CFU,重组率在95%左右,插入片断大小为0.5～4.0kb,多数在1.0kb以上。随机测序72条cDNA,可得到有功能注释的37条全长cDNA和18条编码未知蛋白的基因序列。通过Gene Ontology功能分类可将有功能注释的37个基因分为蛋白质合成、细胞骨架、细胞信号传导、代谢、转运、能量、转录、抗病及防御、生殖发育和未知功能基因等10类,其中蛋白质合成类基因最多(27.03%),与细胞骨架(13.51%)、细胞信号传导(13.51%)及代谢类基因(13.51%)共占67.56%。【结论】构建凡纳滨对虾肌肉组织的全长cDNA文库,可实现短期内获得大量凡纳滨对虾肌肉组织的功能基因表达信息。[Objective]The studies had been undertaken in order to understand the biological basis of expression of functional genes in muscular tissue of Litopenaeus vannamei. [Method]The full length cDNA library from muscular tissue of Litopenaeus vannmei was constructed and expressed Sequence Tags （ESTs） were sequenced. [Result]The constructed library was 8.50×106 CFU in capacity with 95% recombinant coefficient. The PCR results showed that the inserts ranged from 0.5 to 4.0 kb and most of them were larger than 1.0 kb. 72 clones were randomly selected and sequenced for full length. Of which, 37 cDNA sequences were identified with known functions, and 18 cDNA sequences remained as uniden- tified. Using gene ontology function classification, 37 cDNA sequences with known function were classified into groups of protein synthesis, cytoskeleton, signal transduction, metabolism, transporter, energy, transcription factors, response to disease/defense, reproduction development and unknown function ,respectively. The protein synthesis-related genes were the highest in number （27.03%）, followed by cytoskeleton （13.51%）, signal transduction （13.51%）, and metabolism （13.51%）, which collectively accounted for 67.56% of the total identified genes. [Conclusion]The information in constructed full length cDNA library of Litopenaeus vannmei muscular tissue can be used in further studies related to the expression and function of genes in Litopenaeus vannmei.

关 键 词：凡纳滨对虾 肌肉组织 cDNA全长文库 生长性状 

分 类 号：S945.1[农业科学—水产养殖]