摘要
为给研究GlnR蛋白的功能奠定基础,将广西患病罗非鱼无乳链球菌glnR基因克隆并目的基因进行原核表达,根据GeneBank数据库中无乳链球菌基因组序列设计引物,扩增了罗非鱼源无乳链球菌的glnR基因,并通过pGEX-4T-3载体在大肠杆菌中进行原核表达。结果表明:扩增glnR基因的ORF全长372bp,IPTG诱导表达的最佳浓度为3.0mmol/L,最佳诱导时间为5h;在37℃下诱导,重组蛋白大部分以可溶性蛋白的形式存在。
In order to lay the foundation for studying on protein GlnR, the glnR gene of diseased tilapia S. agalactia from Guangxi was cloned and the prokaryotic expression of target gene was conducted. According to the glnR gene sequence of S. agalactiae published in GeneBank, the glnR gene of S. agalactiae from tilapia was amplified by PCR. Then the target gene was expressed in E. coli JM109 through the vector pGEX-4T-3. The results showed that the ORF of gln R gene was 372 bp, the optimal concentration of IPTG was 3.0 mmol/L and the best induction time was five hours. The fusion protein induced in 37℃ mostly existed in the form of soluble protein.
出处
《贵州农业科学》
CAS
北大核心
2013年第5期20-23,共4页
Guizhou Agricultural Sciences
基金
现代农业产业技术体系专项资金项目"国家罗非鱼产业技术体系"(CARS-49)
广西科技重大专项项目"罗非鱼规模化养殖生态工程研究与示范"(桂科攻1123006-3)
关键词
罗非鱼
无乳链球菌
glnR基因
原核表达
tilapia
Streptococcus agalactiae
glnR gene
prokaryotic expression
