应用PCR技术定向引入DNA小片段的策略  

The Strategy of Directed Introducted Low Molecular Weight DNA by PCR

作　　者：张传生[1] 耿立英[2] 孟春花[2] 杨娜娜[2] 朱靖[2] 杜立新[3] 

机构地区：[1]山东农业大学生命科学学院,泰安271018 [2]山东农业大学动物科技学院,山东271018 [3]中国农业科学院畜牧研究所,北京100094

出　　处：《生物技术通报》2005年第3期34-36,共3页Biotechnology Bulletin

摘　　要：描述一种应用PCR技术定向引入DNA小片段和特异酶切位点的方法。为了获得m2/loxp66EGFPloxp71基因片段。根据EGFP基因序列,设计一对特异引物,上、下游引物分别引入m2/loxp66、loxp71序列和Xhol、Mlu1酶切位点。以pEGFPN1质粒为模板,采用PCR扩增以合成DNA双链,插入到克隆载体pMD18T。对重组子测序结果表明,实现了DNA小片段和酶切位点的定向引入。A method of directed introduced low DNA fragments and site during construction of recombinant vector was discried .Using pEGFP-N1 plasmid as the template ,the gene EGFP was amplified by polymerase chain reaction with a pair of specific containing the sites of Xhol,Mlu1 and these sequences m2/loxp66,loxp71. Then the m2/loxp66-EGFP-loxp71 fragment was subcloned into cloning vector pMD-18T .The recombinant plasmid was identified by sequencing. The results showed that the sites and low molecular weight DNA were successfully introduced .

关 键 词：DNA小片段 PCR技术 DNA重组 RMCE基因打靶载体 

分 类 号：Q78[生物学—分子生物学]