重叠延伸PCR合成血管内皮生长因子基因  

Synthesis of the VEGF Gene by Overlap PCR Method

作　　者：张继红[1] 张靖[1] 王俐[1] 杨保胜[1] 王天云[1] 

机构地区：[1]新乡医学院,河南新乡453000

出　　处：《生物技术》2010年第5期40-42,共3页Biotechnology

基　　金：河南省科技攻关项目(0624410041)资助

摘　　要：目的:重叠延伸PCR(OE-PCR)方法合成人血管内皮生长因子(VEGF)基因。方法:根据Genbank中人VEGF基因的编码序列合成15个寡核苷酸片段,相邻引物间均有19个碱基互补,经过7轮PCR循环扩增出人VEGF基因,并将其克隆至pMD18-Tvector,挑取3个重组质粒进行筛选并测序。结果:酶切和测序鉴定结果表明,其中一个与VEGF的同源性达到100%。结论:动态模板OE-PCR基因合成法成功获得576bp的VEGF全长DNA。Objective：To synthesize the human vascular endothelial growth factor （VEGF） by overlap extension PCR （OE-PCR）.Method：According to the VEGF coding sequence in Genbank,fifteen single-stranded oligonucleotides were synthesized ,overlaps of 19 bases between adjacent oligonucleotides.The full-length VEGF gene was amplified directly through seven PCR cycles,then was cloned into the pMD18-T vector.Three recombinant plasmids were selected and sequenced.Result：The results showed that one of the VEGF homology with 100%.Conclusion：We obtained 576bp of VEGF full-length successfully by OE-PCR gene synthesis method.

关 键 词：血管内皮生长因子 重叠延伸PCR 动态模板 基因合成 

分 类 号：Q781[生物学—分子生物学]