35S启动子定量PCR非同源竞争模板的构建

Generation of non-homologous competitor of 35S promoter for QC-PCR

利用35S启动子特异引物,以大肠杆菌基因组DNA为异源模板,经低严谨度PCR扩增并回收克隆适宜DNA片段,构建了35S启动子的非同源竞争对照。测序结果表明,该片段与35S启动子相应序列即靶序列除两端引物序列完全相同外,其他部分没有同源性。利用该非同源竞争对照,建立了转基因玉米35S启动子的QC-PCR技术体系,并进行了校正。

In this article, the E. coli genomic DNA was low stringently amplified by cross - species PCR with the special primers for 35S promoter. The suitable DNA fragment was reclaimed and cloned as a non-homologous competitor. This competitor had no homology with the corresponding sequence of 35S promoter other than the primers at its ends. With this competitor the QC - PCR system for genetically modified maize was established and calibrated.