大肠杆菌glgC基因的克隆及原核表达

以大肠杆菌BL21染色体DNA为模板,根据glgC基因的全序列设计了1对引物,在优化的PCR反应条件下扩增出了glgC基因片段,测序结果显示该片段大小为1296 bp,编码432个氨基酸残基。将该基因克隆到原核表达载体pET-28a-c( )中,重组载体pET-glgC转化至大肠杆菌BL21(DE3),经IPTG诱导后,SDS-PAGE电泳鉴定,得到了与理论推算的glgC基因表达产物分子质量(约53 kD)相符的特异蛋白条带。

Cloning and Prokaryotic Expression of glgC Gene of E. coli

A pair of primers were designed by using genomic DNA of E.coli BL21 as the template and according to the whole sequence of glgC gene,then a glgC fragments were amplified with the pair of primers under optimized PCR conditions and the sequencing of the fragment indicated that the fragment had a size of 1 296 bp and encoded 432 amino acid residues. This gene was cloned to prokaryotic vector pET-28a-c ( ) and then recombinant vector pET- glgC was transformed into the host cells of E. coli BL21 (DE3) ;the host cells were induced with IPTG and then identified by SDS-PAGE to have a specific protein whose molecular weight was the same as the theoretically-calculated one (about 53 kD) of the expression product of glgC gene.