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目的对pGEX-4T-1/gG-2重组质粒进行诱导表达,并纯化、鉴定表达的目的蛋白。方法用最佳诱导条件对重组质粒进行诱导表达,表达产物经SDS-PAGE电泳检测验证后,对存在于超声离心上清液中的融合蛋白用亲和层析技术进行纯化,并用ELISA间接法对纯化的GST/gG-2融合蛋白的灵敏性和特异性等生物活性进行鉴定。结果目的蛋白经诱导后表达于上清中,经鉴定纯化的目的蛋白保留了天然蛋白原有的生物活性。结论 GST/gG-2融合蛋白的获得,为研制以重组蛋白代替传统的全病毒作为检测抗原的新型HSV-2型特异性免疫学检测试剂盒奠定了基础。
Objective To express pGEX-4T-1 / gG-2 recombinant plasmid and purify it, and to identify the expressed protein. Methods The recombinant plasmid was induced by the best induction conditions. The expressed product was verified by SDS-PAGE electrophoresis. The fusion protein in the ultrasonic centrifugal supernatant was purified by affinity chromatography and identified by ELISA indirect method The sensitivity and specificity of the purified GST / gG-2 fusion protein and other biological activities were identified. Results The target protein was expressed in the supernatant after induction. The purified target protein retained the original biological activity of the native protein. Conclusion The obtainment of GST / gG-2 fusion protein lays a foundation for the development of a new HSV-2 type specific immunological detection kit that replaces the traditional whole virus as the detection antigen.