摘要
通过对扩增的金黄色葡萄球菌sep基因序列进行载体构建和双酶切鉴定,将酶切验证的pET-30a-SEP重组质粒分别转入不同的大肠杆菌表达宿主中,选择出最佳表达宿主菌,并进一步优化表达条件,实现重组融合蛋白(rSEP)的可溶性表达;采用Ni^(2+)亲和层析最终获得纯化的rSEP蛋白.研究结果为后续制备单抗、诊断试剂及肠毒素SEP蛋白的致病机制研究奠定基础.
In this study,sep gene from Staphylococcus aureus was constructed into plasmid pET-30a,which was validated by Nde I/Xho I digestion. Then,the recombinant expression plasmid pET-30a-SEP was transformed into different competent cells of E. coli host. The best expression conditions were optimized. Based on that,the soluble expression of recombinant fusion protein（ rSEP） was realized. Further,the soluble rSEP protein was purified by Ni^（2＋） affinity chromatography. The results laid a good foundation for further study of monoclonal antibodies preparation,diagnostic reagents,and pathogenic mechanism of staphylococcal enterotoxin P protein.
作者
唐俊妮
罗双华
刘骥
赵燕英
陈娟
汤承
TANG Jun-ni;LUO Shuang-hua;LIU Ji;ZHAO Yan-ying;CHEN Juan;TANG Cheng(School of Life Science and Technology,Southwest Minzu University,Chengdu 610041,P.R.C.)
出处
《西南民族大学学报（自然科学版）》
CAS
2018年第5期455-461,共7页
Journal of Southwest Minzu University(Natural Science Edition)
基金
国家重点研发计划(2018YFD0500500)
中央高校基本科研业务费重点项目(2018NZD14)
关键词
葡萄球菌新型肠毒素SEP
载体构建
表达条件优化
蛋白纯化
staphylococcal new identified enterotoxin SEP
expression vector construction
expression optimization
protein purification
