白颖苔草热激转录因子(HSF1)真核表达载体的构建

Construction of eukaryotic expression vector bearing the heat shock factor in Carex rigescens

根据表达载体PBI 121特性及酶切位点设计合适的引物,由表达引物通过PCR技术从含有白颖苔草(Carex rigescens)CrHsf全长cDNA的克隆载体的大肠杆菌上扩增出带有特定酶切位点的CrHsf完整开放阅读框。再对载体和目的片段进行酶切处理,处理后将正确的目的基因片段亚克隆至PBI 121植物表达载体。通过PCR及酶切鉴定,结果证明,目的基因片段已被正确克隆到表达载体上,载体构建成功。

Based on the characteristics of the expression vector PBI 121,and its enzyme sites,the appropriate primers were designed.Using the expression primers,we amplified the Escherichia coli bearing the cDNA with CrHsf of Rigens Sedge,and the open reading frame containing the right enzyme sites of CrHsf was got.The expression vectors and the aimed fragments were digested respectively,and then,the aimed DNA fragments were cloned into PBI 121 vector.Through PCR,double enzyme restriction analysis,it was confirmed that the PBI 121/HSF recombinant plasmid was successfully constructed.