GST-HRB融合蛋白的表达与纯化  

The Expression and Purification of GST-HRB Fusion Protein

作　　者：刘丽杰[1] 高学娟[1] 刘小会[1] 陈妙娟[1] 朱森[1] 刘朗夏[1] 

机构地区：[1]暨南大学生命科学技术学院,广州510632

出　　处：《生物技术通报》2011年第11期172-176,共5页Biotechnology Bulletin

基　　金：广东省自然科学基金项目(9151065004000005);中央高校基本科研业务费专项资金资助项目(21609705;21609317)

摘　　要：构建GST-HRB重组质粒,进行融合蛋白的表达、纯化及鉴定。利用PCR扩增及基因重组技术,以pcDNA-3.1-HRB为模板扩增出HRB全基因序列,并将其插入带有GST(谷胱甘肽巯基转移酶)标签的原核表达载体pGEX-6P-1中,构建GST-HRB融合蛋白表达质粒。然后,将重组质粒GST-HRB转化至大肠杆菌Rosseta进行融合蛋白的表达。利用GST琼脂糖珠进行融合蛋白的纯化,最后应用SDS-PAGE电泳和Western blotting鉴定纯化的融合蛋白。结果表明,成功构建pGEX-6P-1-HRB原核表达载体,表达及纯化了GST-HRB融合蛋白。It was to construct a GST-HRB recombinant plasmid,and identify fusion protein.Using PCR and recombinant DNA technology,HRB full gene sequence was amplified from template pcDNA-3.1-HRB and inserted into the prokaryotic expression vector pGEX-6P-1 with tag GST（glutathione sulfurtransferase）.The recombinant vector was then transformed into the E.coli Rosetta strain to produce the GST-HRB fusion protein,which was purified through a Sepharose-Glutathione column,and verified by SDS-PAGE and Western blotting using an anti-GST polyclonal antibody.We successfully constructed the GST-HRB plasmid,produced and purified the GST-HRB fusion protein,which would lay a foundation for further research of HRB protein.

关 键 词：GST-HRB 构建 融合蛋白 表达 纯化 

分 类 号：Q78[生物学—分子生物学]