摘要
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。
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 1296 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.
出处
《西北植物学报》
CAS
CSCD
北大核心
2006年第4期667-671,共5页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家863计划项目(2004AA241132)甘肃省科技攻关项目(2GS054-A41-005-01)
关键词
glgC基因
克隆
原核表达
载体构建
glgC gene
cloning
prokaryotic expression
vector construction
