RT-PCR克隆广谱抗病基因NPR1及其蛋白表达载体构建

Cloning of Broad-spectrum Anti-disease Gene NPR1 with RT-PCR and Construction of Its Protein Expression Vector

[目的]克隆植物广谱抗病基因NPR1并构建其蛋白表达载体。[方法]提取拟南芥总RNA,设计相关引物,采用反转录PCR方法克隆NPR1基因;利用酶切连接方法,将该基因正向导入蛋白表达载体。[结果]经过相关检验,将NPR1正向插入pMXB10载体中,得到了pMXB10-NPR1蛋白表达载体。[结论]成功构建了包含NPR1的蛋白表达载体。

[Objective]It is to clone broad-spectrum anti-disease gene NPR1 and to construct its protein expression vector.[Method]First extract Arabidopsis thaliana total RNA and design relevant primers,and then the method of reverse transcription PCR is adopted.With the method of enzyme ligation,this gene will be directed into protein expression vector.[Result]After relevant testing,NPR1 will be inserted into vector pMXB10 to obtain pMXB10-NPR1 protein expression vector.[Conclusion]Protein expression vector with NPR1 will be successfully constructed.