家猪百日胚胎胸腺的基因表达谱  

Gene expression pattern in 100-day porcine foetus thymus

作　　者：陈艳炯[1] 邓亚军[1] 叶琳[2] 耿佳宁[3] 李生斌[1] 

机构地区：[1]西安交通大学医学院法医系,陕西西安710061 [2]杭州华大基因研究中心,浙江杭州310007 [3]北京华大基因研究中心,北京101300

出　　处：《第四军医大学学报》2003年第20期1837-1840,共4页Journal of the Fourth Military Medical University

基　　金：中国丹麦合作家猪基因组计划 (2 0 0 2AA2 2 90 61 )

摘　　要：目的 :构建家猪 1 0 0d胚胎胸腺基因表达谱 ,为研究胸腺的发育和分化机制奠定基础 .方法 :随机挑取家猪1 0 0d胸腺cDNA文库 (FTY)的 1 1 71 2个克隆 ,进行单向全自动大规模测序 ,去污染后共获得 70 71个高质量的表达序列标签 (EST)用于基因表达分析 .结果 :对 70 71个高质量EST的聚类分析结果获得 95 9个多拷贝基因和 30 74个单拷贝基因 .与非冗余的核酸数据库、Swiss Prot数据库及人、牛和鼠的U nigene数据库进行BLAST ,发现 80 6个多拷贝基因和 1 6 6 9个单拷贝基因 (共计 5 4 4 2个ESTs)与已知序列有同源性 ,1 6 2 9个EST没有发现同源序列被认为是新基因 .根据功能将已知基因归为 7类 ,其中基因表达相关基因占 36 .99% ,胸腺特异的防御功能相关基因有一定程度表达 (占 7.5 5 % ) ,这与人婴儿胸腺基因表达谱相类似而不同于成年人胸腺组织 .将 1 0 9个既发现有ORF又具有PolyA的多拷贝新基因与人类基因组框架序列进行BLAST ,发现其中 5 8个有同源序列 .结论 :EST技术是发现新基因的有效手段之一 ;此期家猪胸腺仍处于发育阶段 .AIM: To get a better understanding of the thymus development and differentiation,we constructed the gene expression pattern of 100 day foetus porcine thymus(FTY). METHODS: 11712 randomly picked clones from FTY cDNA library were single pass sequenced. After contamination was removed, 7071 cleaned ESTs were used for gene expression analysis. RESULTS: After clustering the 7071 cleaned ESTs by PHRAP, 959 contigs(size>1) and 3074 singletons were obtained. Blast search showed that 806 contigs and 1669 singletons (totally 5442 ESTs) had homologues in nonredundant nucleic acid database, the Swiss Prot database and Unigene database of human, cattle and mouse. The 1629 ESTs without similarities were considered novel. According to function, the known genes were classified into 7 group and 36.99% of ESTs were classified into the gene expression group. The thymus functional genes were expressed in a certain degree (accounting for 7.55%). Comparative expression analysis showed that the gene expression pattern of FTY was very similar to human infant thymus gene expression profile and different from that of adult human. Blast search found that 58 of 109 novel sequences with both poly A and ORF prediction had homologues to human genome sequence. CONCLUSION: EST strategy is economical and virtual in finding new genes. The 100 day porcine thymus is still at a developmental stage.

关 键 词：猪 胸腺/胚胎学 EST 

分 类 号：Q456[生物学—生理学] Q343.1