胰岛素前体基因克隆及其原核表达载体的构建

Insulin Precursor Gene Cloning and Prokaryotic Expression Vector Construction

目的:构建胰岛素前体基因的原核表达载体。方法:以重叠PCR方法扩增获得胰岛素前体基因片段,并将片段克隆入zero pCR4 Blunt-TOPO载体内,获得胰岛素前体中间载体;随后将胰岛素前体克隆到Pet23表达载体中。结果:双酶切鉴定结果显示,成功构建了胰岛素前体基因的原核表达载体。结论:本实验成功构建了胰岛素前体基因的原核表达载体,为后续实现大规模的实验室表达和胰岛素的工业化生产奠定了坚实的基础。

Objective：TO construct the prokaryotic expression vector of insulin precursor gene. Method：Insulin precursor gene fragment was amplified by overlap PCR,and cloned into zero pCR4 Blunt-TOPO load in vivo,to obtain insulin precursor intermediate vector. The insulin precursor was cloned into Pet23 expression vector. Results：The result of enzyme digestion showed the prokaryotic expression vector of insulin precursor gene was successfully constructed. Conclusion：We successfully construct the prokaryotic expression vector of insulin precursor gene,this lays a solid foundation for the large-scale laboratory expression and industrialized production of insulin.