热诱导的位点特异性重组植物表达载体构建及烟草转化

Construction of Plant Expression Vector with Heat Shock Induced CinH /Rs Site-specific Recombination System and Transformation of Nicotiana Tabacum

选择标记基因安全性问题是限制转基因植物商品化种植的重要因素。为了从转基因后代中诱导性剔除选择标记基因,本研究克隆了拟南芥热激蛋白Hsp18.2基因启动子,并构建了Hsp18.2启动子诱导表达的CinH/Rs位点特异性重组删除植物表达载体。结果表明,利用农杆菌介导法进行了烟草转化,转化后的烟草经过除草剂筛选和PCR检测,得到转基因阳性植株。本研究为利用位点特异性重组建立植物无标记基因转化体系奠定了基础。

The security problem of selectable marker genes has become one of the important factors limiting the commercial plantation of transgenic plants.In order to delete the marker genes from transgenic plants,in this paper,the promoter of Hsp18.2 heat shocked protein gene was cloned from Arabidopsis,and the plant expression vector of heat shock induced CinH/Rs site-specific recombination system was constructed.The result showed that after transformation of nicotiana tabacum mediated by Agrobactrium tumefaciens,the transgenic plants were screened by PPT selection and PCR analysis to get transgenic positive genes.This study would lay an important foundation for the marker free plant transformation using site-specific recombination system.