密码子优化提高重组金葡菌肠毒素O在大肠杆菌中的表达水平  

Codon optimization of recombinant staphylococcal enterotoxin O enhances the expression level in Escherichia coli

作　　者：黄鹏[1] 孙红颖[1] 陈枢青[1] 

机构地区：[1]浙江大学药学院药物毒理与生化药学研究所,浙江杭州310058

出　　处：《浙江大学学报（医学版）》2011年第3期297-303,共7页Journal of Zhejiang University(Medical Sciences)

摘　　要：目的:优化稀有密码子提高重组肠毒素O的表达量。方法:将带H is标签的肠毒素O成熟肽克隆至pET28a质粒上,采用PCR方法对15个稀有密码子进行突变,将突变后的重组质粒转化入大肠杆菌BL21(DE3),并进行蛋白表达及活性鉴定。结果:通过稀有密码子优化,重组肠毒素O的表达量由菌液总蛋白的7.49%提高至19.8%;小鼠淋巴细胞增殖试验表明,通过密码子优化的肠毒素O具有刺激淋巴细胞增殖的作用,并且其增殖作用与未优化前的肠毒素O相当。结论:稀有密码子优化能够有效增加肠毒素O的表达量。Objective:To enhance the expression level of staphylococcal enterotoxin O(SEO) by optimization of rare codons.Methods:The gene of mature SEO(His-tag included) was cloned to pET28a,and 15 rare codons on the gene were optimized by PCR technology.These recombinant plasmids then were transformed into E.coli BL21(DE3),respectively.After IPTG induced,the expression levels of those mutants were analyzed by SDS-PAGE.The proteins were purified and their bioactivities were determined.Results:After the optimization of rare codons,the expression levels were increased from 7.49% to 19.8% in total cell proteins.The optimized SEO had bioactivity to stimulate the proliferation of murine lymphocytes,which was equivalent to that of non-optimized SEO in vitro.Conclusion:Optimization of rare codons can enhance the expression of SEO effectually.

关 键 词：葡萄球菌 金黄色/免疫学 肠毒素类/免疫学 密码子 基因表达 遗传载体 质粒 突变 细胞增殖 淋巴细胞 

分 类 号：Q93[生物学—微生物学]