Cre-lox重组系统介导转基因烟草中外源基因删除的研究  

Cre/loxP-mediated Deletion of Transgene in Transgenic Tobacco

作　　者：甄伟[1] 汪阳明 丁伟[1] 陈溪[1] 胡鸢雷[1] 林忠平[1] 

机构地区：[1]北京大学蛋白质及植物基因工程国家重点实验室

出　　处：《北京大学学报（自然科学版）》2001年第4期477-482,共6页Acta Scientiarum Naturalium Universitatis Pekinensis

基　　金：高等学校博士学科点专项科研基金资助项目

摘　　要：对Cre在转基因个体中介导的重组效率进行了研究。构建了含有Cre基因 (p35S Cre)和GUS基因侧翼含同向loxP位点的 (loxP p35S GUS loxP)两种植物表达载体。以共转化的技术将两种基因元件同时转化烟草得到转基因植株 ,根据对共转化植株GUS基因的活性分析、分子检测、PCR检测及对重组后扩增DNA片段进行序列分析表明 :Cre loxP重组系统在转基因烟草中能精确高效地介导转基因的删除 ,但也存在部分植株不能完全删除的现象。In the present study,we develop an efficient method to be used to examine recombination efficiency of transgenic plantlet.Two constructs,one containing Cre recombinase under the control of 35 S promoter (p35S/Cre) and the other containing a \%uidA\% gene driven by 35 S promoter flanked by loxP site (loxP/35S/GUS/loxP),were simultaneously introduced into tobacco by cotransformation method.The results show that Cre loxP system can precisely and efficiently direct deletion of transgene based on Gus assay,PCR analysis and sequence of recombination DNA fragment,but also find that incomplete deletion exist in partial plant.

关 键 词：Cre-loxP重组系统 转基因烟草 共转化 GUS活性分析 不完全删除 重组效率 LOXP位点 

分 类 号：Q784[生物学—分子生物学]