人SMYD3基因原核表达载体的构建及在大肠杆菌中的表达

构建人SMYD3基因原核表达载体,并在大肠杆菌中诱导表达。通过PCR方法从重组质粒pcD-NA-SMYD3中扩增得到SMYD3基因,将其克隆入pGEX-6P-1原核表达载体。将重组质粒转化入E.coli Rosetta(DE3)中,以IPTG诱导融合蛋白表达。表达产物用SDS-PAGE及Western blotting分析鉴定。酶切鉴定和测序结果证明成功构建了重组表达载体pGEX-SMYD3。SDS-PAGE及Western blotting鉴定结果表明人SMYD3基因在大肠杆菌中获得了良好的表达。实验成功地构建了SMYD3基因原核表达载体并在大肠杆菌中获得良好表达,为进一步研究SMYD3蛋白功能奠定了基础。

Construction and Expression of Human SMYD3 Gene in E. coli Rosetta (DE3)

To construct and to express of human SMYD3 gene in E.coli.The SMYD3 gene was obtained from pcDNA-SMYD3 vector by PCR and then cloned into prokaryotic expression vector pGEX-6P-1.The recombinant plasmids were transformed into E.coli Rosetta(DE3).GST-SMYD3 fusion protein was induced to express in E.coli Rosetta(DE3) and identified by SDS-PAGE and Western blotting.The recombinant vector was constructed and expressed in E.coli successfully.The study has laid a foundation for further study on the biological function of human SMYD3 gene.