鸭早期胚胎发育相关基因消减文库的构建与分析  

Construction and Analysis of a Subtractive cDNA Library of Duck Early Embryonic Development Related Genes

作　　者：钟灵秀[1,2] 刘雅丽[1,2] 李进军[1] 沈军达[1] 卢立志[1] 石放雄[2] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,杭州310021 [2]南京农业大学动物科技学院,南京210095

出　　处：《农业生物技术学报》2011年第4期763-770,共8页Journal of Agricultural Biotechnology

基　　金：农业部现代农业产业技术体系建设专项资金(No.NYCYTX-45-02);浙江省重大科技攻关项目(No.2005C12005-01;No.2008C14073)共同资助

摘　　要：动物胚胎发育受到多个基因的表达调控,为探索鸭早期胚胎发育的分子机制,本实验构建了鸭早期胚胎发育相关基因的消减文库,筛选与鸭早期胚胎发育相关的候选基因。采集两个不同时段的鸭(Anas platy rhynchos)胚(6和20h)分别作为驱动组(driver)和实验组(tester),通过由双链特异性核酸酶(designated duplexspecific nuclease,DSN)介导的消减杂交方法,构建鸭早期胚胎发育相关基因的消减文库,并进行生物信息学分析。结果表明,在构建好的文库中随机挑选216个阳性克隆进行测序,最终共得到54条有效序列,平均长度为1084bp。将这些序列在GenBank中进行BLAST比对,其中44条有同源基因,基因注释(gene ontology,GO)显示,这些基因与代谢、建立细胞定位、生物调节等各种生物途径相关,它们在鸭早期胚胎发育过程中的不同方面(如神经系统发育、免疫调节、转录调节等)都发挥着非常重要的作用。这些差异表达基因的分离与鉴定,有助于家禽早期胚胎发育的相关研究。Animal embryonic development is regulated by many genes.In order to explore the molecular mechanism of duck early embryonic development,we constructed the subtractive library of duck early embryonic development related genes to screen the related candidate genes of duck early embryonic development.Two different periods（6 and 20 h） duck（Anas platyrhynchos） embryo were used respectively as the driver and tester,and a designated duplex-specific nuclease（DSN） mediated subtractive hybridization method was employed to construct the subtractive library of duck early embryonic development related genes,and the genes were sequenced and analyzed by bioinformatics.Results showed that 54 genes were obtained by DNA sequencing from 216 positive clones picked randomly.Their average length was 1 084 bp.Nucleotide BLAST homological analysis showed that 44 genes had similarities to known genes.Gene ontology showed that these genes were related with a variety of biological pathways,such as metabolism,establishment of cellular localization,biological regulation,etc.And these genes played very important roles during early embryonic development of duck in many different aspects（such as nervous system development,immune regulation,transcriptional regulation,etc.）.We conclude that isolation and identification of these differentially expressed genes will contribute to the future related research of poultry early embryonic development.

关 键 词：鸭 早期胚胎发育 双链特异性核酸酶(DSN) 消减杂交 

分 类 号：Q954.4[生物学—动物学]