广西医科大学学报2017，Vol.34Issue(12)：1686-1690,5.DOI:10.16190/j.cnki.45-1211/r.2017.12.002
齿垢密螺旋体精氨酸激酶TDE2037的原核表达与纯化
Prokaryotic expression and purification of an arginine kinase gene TDE2037 of Treponema denticola
摘要
Abstract
Objective:To construct the prokaryotic expression vectors of Treponema denticola gene TDE2037,and express and purify.Methods:TDE2037 was amplified by PCR and inserted into pET-21a and pET28a-SU-MO plasmids.Recombination plasmids were transformed into E.coli BL21 to express recombined protein under induction of IPTG.The expressed products were identified by biological mass spectrometry and the targeted protein was purified with metal affinity chromatography column.Results:TDE2037 was amplified successfully and cloned into pET-21a and pET28a-SUMO plasmids.Sequences were identified to de consistent with the data of Genbank.Recombination proteins were successfully expressed in E.coli BL21.The protein of pET-21a-TDE2037 completely existed in form of inclusion body and remained insoluble,whereas the expression product of pET28a-TDE2037-SUMO,partially soluble and partially in form of inclusion body.High purity protein was achieved by metal affinity chromatography column.Conclusion:The prokaryotic expression vectors of Treponema denticola gene TDE2037 were successfully constructed and the expressed protein was successfully purified,which may lay a basis for the further study of TDE2037.关键词
齿垢密螺旋体/蛋白纯化/精氨酸激酶Key words
Treponema denticola/protein purification/arginine kinase分类
生物科学引用本文复制引用
陈宇星,周鹏,唐路,陈文霞..齿垢密螺旋体精氨酸激酶TDE2037的原核表达与纯化[J].广西医科大学学报,2017,34(12):1686-1690,5.基金项目
国家自然科学基金资助项目(No.81160133) （No.81160133）
