小拟南芥chitinase基因原核表达载体的构建及其在大肠杆菌中的表达

Construction of prokaryotic expression vector of Arabidopsis pumila chitinase and its expression in Escherichia coil

以野生资源小拟南芥(Arabidopsis pum ila)chitinase基因的cDNA为基础,采用基因重组技术,将该基因按正确的阅读框架定向克隆于原核表达载体pET-30a(+)中,转化大肠杆菌BL21(DE3),用IPTG诱导表达,并对表达产物进行SDS—PAGE分析。结果表明:重组小拟南芥chiti-nase基因在大肠杆菌中获得高效表达,其分子量约为40 KD。小拟南芥chitinase基因原核表达载体的成功构建和重组小拟南芥chitinase蛋白在大肠杆菌中的高效表达,为进一步研究其生物学功能奠定了基础。

Prokaryotic expression vector pET-CH of Arabidopsis pumila chitinase gene cDNA was construtted by means of recombinant gene technology and expressed in Escherichia coil BL21 under induction of IPTG, the expressed products were detected by SDS-PAGE analysis. Result show： Recombinant chitinase was efficiently expressed in E. coli BL21, it molecular weight was about 40 KD. The successful construction of prokaryotic, expression vector containing Arabidopsis pumila chitinase gene and effective expression of recombinant chitinase in E. coli BL21 laid the foundation for further study on its biological function.