论文部分内容阅读
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。
Purpose: Recombinant fusion protein expressing the RV specific segment of RV. Methods: The positive recombinant plasmid vector pGEX-2T / E1-N identified by double enzyme digestion and sequencing was transformed into competent E. coli BL21 and induced with isopropyl-β-D thiogalactoside (IPTG) Expression, and optimization of induction conditions. Recombinant fusion protein was purified with glutathione sepharose beads and identified by SDS-PAGE. Results: The recombinant fusion protein was induced by IPTG. The optimal inducer concentration was 1 mmol / L at 37 ℃ and the optimal induction time was 4 h. The recombinant fusion protein was expressed in soluble form when the temperature was lowered from 37 ℃ to 16 ℃. Purified recombinant fusion protein was obtained by using glutathione sepharose beads. Conclusion: The recombinant fusion protein of purified specific fragment of rubella virus E1 can be obtained by using prokaryotic expression system.