拟南芥CGP1基因启动子克隆及其GUS转基因植株的构建

Cloning of CGP1 gene promoter and construction of GUS transgenic plants in Arabidopsis thaliana

前期研究发现一个功能未知基因响应植物镉胁迫,将之命名为CGP1。文章为探究植物基因CGP1的表达模式及其在响应镉胁迫中的表达变化情况,以野生型拟南芥为实验材料构建CGP1基因启动子接pART7-GUS载体及其转基因植株。通过提取野生型拟南芥的DNA,并以DNA为模板利用聚合酶链式反应(polymerase chain reaction, PCR)技术克隆得到CGP1基因启动子序列,将启动子序列和GUS载体双酶切后进行连接,并转化到大肠杆菌感受态细胞中,通过测序获得构建成功的ProCGP1-GUS载体。利用浸染花序法将ProCGP1-GUS载体转入野生型拟南芥中,并通过抗性筛选获得ProCGP1-GUS转基因阳性植株,为研究镉胁迫下CGP1基因的功能奠定了基础。

Previous studies have found that a gene with unknown function responds to cadmium stress in plants, named as CGP1. In order to further study the function of plant gene CGP1 and its expression pattern in response to cadmium stress, wild-type Arabidopsis thaliana was used as experimental materials to construct CGP1 gene promoter with pART7-GUS vector and transgenic plants. The DNA of wild-type Arabidopsis thaliana was extracted and used as a template to clone the promoter sequence of CGP1 gene by using polymerase chain reaction(PCR) technology. The promoter sequence was coupled with GUS vector after double digestion, and then transformed into Escherichia coli competent cells. The ProCGP1-GUS vector was successfully constructed by sequencing. It was transformed into wild-type Arabidopsis thaliana by the inflorescence method, and the ProCGP1-GUS transgenic positive plants were obtained through resistance screening, which laid the foundation for studying the function of CGP1 gene under cadmium stress.