摘要
采用大肠杆菌表达体系来获得C-反应蛋白(CRP)融合蛋白。以pDNR-CRP质粒为模板,用PCR方法扩增获得全长的CRP基因,将其克隆入表达载体pET28 a(+)中,再转化到宿主菌BL21(DE3)中,加入IPTG诱导表达,表达产物做SDS-PAGE电泳分析,最后做免疫原性检测。成功地构建了CRP基因原核表达载体pET28 a(+)-CRP,经IPTG诱导SDS-PAGE电泳分析表明C-反应蛋白能在大肠杆菌中获得表达,但免疫原性检测初步结果为阴性。结果证明CRP能以融合蛋白的形式在宿主菌BL21(DE3)中得到大量表达,表达产物缺乏免疫原性,其机理还有待进一步试验探讨。
In this study,the C - reactive protein was prepared by the E. coli expression system. The CRP gene was amplified by PCR with the template of pDNR - CRP plasmid, then cloned into the pET - 28a（ ＋ ）,and transformed into E. coli BL21 （ DE3 ）. The recombinant CRP protein was induced and expressed by isopropylthio- a- D- galactoside（IPTG）. SDS- PAGE analysis showed that the target protein was expressed at a high level in E. coli BL21 （ DE3 ）. However it showed no immunogenicity in the primary detection, further study is needed about its immunogenicity.
出处
《江西农业大学学报》
CAS
CSCD
北大核心
2006年第2期226-229,共4页
Acta Agriculturae Universitatis Jiangxiensis
基金
湖北省教育厅青年基金项目(Q200513002)
国家"863"计划项目(2002AA241031)
关键词
C反应蛋白
原核表达
IPTG
免疫原性
C - reactive protein
prokaryotic - expression
IPTG
immunogenicity
