摘要
目的:表达并纯化M6型GAS表面烯醇化酶以及其敲除C末端赖氨酸残基重组蛋白(rSEN和rSENΔ434-435)。方法:克隆了M6型GAS ATCC32175的SEN基因以及SENΔ434-435基因,与pASK-IBA37载体连接后,表达蛋白并用亲和层析色谱纯化重组蛋白;通过酶促反应实验测定了重组蛋白的生物活性。结果:2种基因克隆条件稳定,蛋白表达量大,且纯化方法可靠,纯化的重组蛋白具有较高的生物活性。结论:成功表达并纯化了rSEN和rSENΔ434-435蛋白。
Objective：To express and purify the recombinant streptococcal surface enolase （rSEN） and its C -terminal lysine residues -truncated variant （rSENA434 -435）. Methods：We cloned sen and sen△434 -435 from M6 -type GAS ATCC32175, produced rSEN and rSEN△434 - 435 in E. coli using the 6 × Histag pASK - IBA37 expression vector and purified the recombinant proteins by af- finity chromatography with TALON metal affinity resins. The enzyme reaction was performed to determine enolase activity. Results： PCR conditions ampifing sen and sen△434 -435 were veridical and expression and purification of recombinant proteins were profuse and convenient. The purified rSEN and rSEN△434 - 435 were found to have relatively full enolase activity. Conclusion ： We succusfully expressed and purified rSEN and rSEN△434 -435.
出处
《内蒙古农业大学学报（自然科学版）》
CAS
北大核心
2011年第1期167-171,共5页
Journal of Inner Mongolia Agricultural University(Natural Science Edition)
基金
国家自然科学基金(30860019)
关键词
A群链球菌
烯醇化酶
表达
纯化
Group A streptococcus
α - enolase
expression
purification
