α2巨球蛋白cDNA片段-增强型绿色荧光蛋白质粒的构建

Construction of alpha-2 macroglobulin cDNA-enhance green fluorescent protein expression plasmid

目的:构建α2巨球蛋白cDNA片段(FP6)-增强型绿色荧光蛋白(pEBFP)质粒(pEBFP/FP6双表达质粒)。方法:利用PCR技术克隆出FP6,将其克隆至pMD18-T载体中,再通过分子克隆技术将FP6插入pEBFP表达基因中,构建pEBFP/FP6质粒,双酶切和测序鉴定。结果:PCR方法能克隆FP6,并成功构建pEBFP/FP6质粒,双酶切和测序结果表明构建的pEBFP/FP6质粒完全符合设计要求。结论:利用PCR、酶切、连接反应等基因克隆手段,可以构建pEBFP/FP6双表达质粒。

Objective：To construct the alpha -2 macroglobulin cDNA （FP6） -enhance green fluorescent protein （pEBFP） expression plasmid（pEBFP/FP6）. Methods： FP6 was generated using PCR and cloned into pMD18 -T vector. The recombinant plasmid of pEBFP/FP6 was constructed using melecular cloning technique, and identified by double enzymolysis and sequencing analysis. Results： FP6 was generated using PCR. The coexpression plasmid of pEBFP/FP6 was constructed and completely consistent with requirement of devise by double enzymolysis and sequencing analysis. Conclusion： pEBFP/FP6 coexpression plasmid can be constructed by gene cloning methods, such as PCR, enzymolysis and coupled reaction.