盐生植物盐穗木HcPS_H基因大肠杆菌表达载体的构建

[目的]构建盐穗木HcPS_H基因的大肠杆菌表达载体。[方法]以质粒pGEM-T-HcPS_H为模板,HcPS_H-pET28a-F、HcPS_H-pET28a-R为引物进行PCR扩增,获得两端含酶切位点的HcPS_H基因目的片段。将该目的片段与大肠杆菌表达载体pET-28a通过T4DNA连接酶连接,并转化大肠杆菌(Escherichia coli)DH5α,得到重组子克隆,抽质粒进行测序鉴定。[结果]扩增出HcPS_H基因片段长度为441 bp,将其连接到大肠杆菌表达载体pET-28a上后,得到重组子质粒,经PCR检测、双酶切验证及测序鉴定,表明成功构建原核表达载体。[结论]该研究成功构建了盐穗木HcPS_H基因的大肠杆菌表达载体,为进一步进行该基因功能的研究奠定了基础。

Constructing of a E.coli Expression Vector of HcPS_H Gene from Halostachys caspica

[Objective]To construct a E.coli expression vector of HcPS_H gene from Halostachys caspica.[Method]The HcPS_H gene full-length fragment was obtained from pGEM-T-HcPS_H by PCR using a pair of including enzyme site primer,and then the purified fragment was cloned into pET-28a vector.[Result]HcPS_H gene was amplified from Halostachys caspica.The gene fragment was 441bp,and was cloned into pET-28a vector.After analysis of restriction endonuclease digestion and PCR,the constructed expression vector was proved to be correct.[Conclusion]The expression vector of HcPS_H gene of Halostachys caspica was constructed successfully.