摘要
用RT-PCR的方法扩增水泡性口炎病毒G蛋白抗原决定基部分基因片段,构建克隆质粒pMD18-T-G660和表达质粒pET-28a-G660,转化宿主菌BL21(DE3),经IPTG诱导后,得到高效表达,表达蛋白质的相对分子质量为30 000,占总蛋白的20.745%。
One DNA fragment of protective antigen genes of vesicular stomatitis virus were amplified by RT-PCR with primers synthesized according to GenBank. The PCR products were cloned into pMD18-T and the recombinant plasmids pMD18-T-G660 was constructed. The connective products were transformed to competent cells of E. coli host strain DH5α. The recombinant plasmid pET-28a-G660 was constructed by cloning pMD18-T-G660 gene into prokaryotic expression vector pET-28a （＋） and expressed in E. coli host cells BL21 （DE3）. The recombinant protein was highly expressed by growing individual clone in normal LB media ,then induced by IPTG. The expressed protein with an apparent MW. of 30 000 was obtained,amounts for 20. 745 percent of the total proteins.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第5期475-477,共3页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30270994)
关键词
抗原决定簇基因
克隆
表达
protective antigen genes
cloning
expression
