Acta Scientiarum Naturalium Universitatis Pekinensis

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Cre/loxP-mediated Deletion of Transgene in Transgenic Tobacco

ZHEN Wei,WANG Yangming,DING Wei,CHEN Xi,Hu Yuanlei,LIN Zhongping   

  1. National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing, 100871
  • Received:2000-04-24 Online:2001-07-20 Published:2001-07-20



  1. 北京大学蛋白质及植物基因工程国家重点实验室,北京,100871

Abstract: 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 35S promoter (p35S/Cre) and the other containing a uidA gene driven by 35S 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.

Key words: Cre/loxP system, transgenic tobacco, cotransformation, GUS assay, incomplete deletion

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。

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

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