摘要
以凡纳滨对虾为研究对象,通过RT-PCR技术扩增出铁蛋白(Ferritin)基因,经特定的酶切(Not1,Sma1)后插入到表达质粒pGEX-4T-2上构建表达载体,重组质粒转化大肠杆菌.经菌落PCR和质粒双酶切鉴定确认,证实成功地构建了对虾铁蛋白基因的原核表达载体.再对重组菌用IPTG诱导表达,采用Glutathione Sepharose 4B亲和层析得到目的蛋白用于制备抗体,为研究铁蛋白在对虾抗病毒免疫中的作用奠定基础.
In this study, ferritin gene from Litopenaeus vannamei was amplified by PCR, digested by Sma I and Not I, and inserted into the expression vector pGEX-4T-2 to construct recombinant plasmid. Colonies with recombinant plasmid were picked for PCR and culture identified by enzyme digestion. The results show that ferritin gene ean be successfully inserted into the pGEX-4T-2 expression vector. After induced with IPTG at 18℃, the fusion protein can be purified with Glutathione Sepharose 4B.
出处
《泉州师范学院学报》
2011年第6期7-10,共4页
Journal of Quanzhou Normal University
基金
国家自然科学基金(31072236)
福建省教育厅项目(JK2009043)
关键词
凡纳滨对虾
铁蛋白
重组
表达
Litopenaeus vannamei
ferritin
recombination
expression
