基因预测与PCR技术相结合从cDNA文库中获取表达基因的研究  

Gene Prediction by GenScan Software Combined with the PCR Technique to Obtain Specific Expressed Genes from cDNA Library

作　　者：谢小燕[1] 谭有将[1] 李景全[1] 陈芳[1] 高剑峰[1] 

机构地区：[1]石河子大学生命科学学院,石河子832003

出　　处：《生物技术通报》2010年第9期61-64,共4页Biotechnology Bulletin

基　　金：中国美利奴绵羊MHC区段基因组成与结构研究项目(国际合作项目;2006DFB33750)

摘　　要：应用计算机工具、GenScan软件预测中国美利奴绵羊MHC Class I区段的BAC文库中453oⅡ克隆的基因数目、特性及结构,建立一种可以从cDNA文库中简便有效获取表达基因的技术方法。选取4个预测基因作研究对象设计引物,应用PCR技术,对已构建好的cDNA文库进行PCR扩增,回收"目的基因"片段并连接pGEM-T载体,转DH5α大肠杆菌中扩增后测序。琼脂糖凝胶电泳检测PCR扩增产物,cDNA文库中有目的条带,测序结果与GenBank进行Blast分析,分析结果表明这些基因与羊的基因均具有99%以上的相似性。因此应用基因预测分析与PCR结合技术可简便迅速的从cDNA文库中获取表达基因。Predicting the number,identity and structure of genes of 453oⅡ cloning,which in the BAC library of Chinese Merino sheep MHC ClassⅠsection by computer and GenScan software,a technical method that could simply and effectively access expressed gene from the cDNA library was established.Four genes as study objections were selected,and specific primers were designed to amplified these genes from the library.PCR produces were linked to the pGEM-T vector,then transform the vector which contain target genes into E.coli DH5α.By agarose gel electrophoresis,we detected the purpose bands,then we sequence the PCR products,blast the sequences with the sheep gene,and the results shows high identity（more than 99%）.Therefore,the technical method that integrated gene predicted analysis and PCR could obtain expressed gene from the library easily and quickly.

关 键 词：基因预测 PCR CDNA文库 

分 类 号：Q78[生物学—分子生物学]