风疹病毒(RV)包膜糖蛋白E1特异基因片段的原核表达

Prokaryotic Expression of Specific Fragment of Rubella Virus E1 Gene

目的:表达风疹病毒(RV)E1特异肽段的重组融合蛋白。方法:经过双酶切鉴定和测序鉴定的阳性重组质粒载体pGEX-2T/E1-N,转化到感受态大肠杆菌BL21后,用异丙基-β-D硫代半乳糖苷(IPTG)诱导其表达,并对诱导条件进行优化。用谷胱甘肽琼脂糖珠纯化重组融合蛋白,SDS-PAGE鉴定。结果:用IPTG可以诱导E1特异肽段的重组融合蛋白表达,37℃诱导时,最佳诱导剂浓度为1mmol/L,最佳诱导时间为4h。诱导温度从37℃降至16℃时,重组融合蛋白以可溶性形式表达,用谷胱甘肽琼脂糖珠纯化获得了纯化的重组融合蛋白。结论:利用原核表达系统可以获得纯化的风疹病毒E1特异肽段的重组融合蛋白。

Objective：To acquire purified recombinant fusion protein of E1-specific peptide of Rubella virus（RV）.Methods：The recombinant plasmid pGEX-2T/E1-N,confirmed by digestion with restriction endonuclease and sequencing,was transferred into E.coli BL21.Fusion protein was induced with isopropylthio-β-D-galactoside（IPTG） and induction conditions were optimized.Recombinant fusion protein rE1-N was purified by glutathione sepharose 4B and identified by SDS-PAGE.Results：Induced by IPTG,recombinant fusion protein was acquired.When induced at 37℃,the optimum concentration of IPTG was 1 mmol/L,and the optimum induction time was 4 h.Induced with lower temperature（transferring from 37℃ to 16℃）,resolvable recombinant fusion protein was expressed.Purified combinant fusion protein was acquired by glutathione sepharose 4B.Conclusion：The purified recombinant fusion protein of E1-specific peptide can be acquired successfully by prokaryotic expression system.