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目的对构建的pGEX-4T-1/gG-1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,对存在于超声离心后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-1融合蛋白进行灵敏性和特异性等生物活性的初步鉴定。结果目的蛋白经诱导后表达于上清中,初步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-1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-1型特异性免疫学检测试剂盒奠定了基础。
OBJECTIVE: To construct recombinant plasmid pGEX-4T-1 / gG-1 and induce the expression of the target protein. Methods The recombinant plasmid was induced by the best induction conditions. The expressed product was verified by SDS-PAGE electrophoresis. The fusion protein present in the supernatant after ultrasonic centrifugation was purified by affinity chromatography and identified by ELISA indirect method Purification of GST / gG-1 fusion protein sensitivity and specificity of biological activity of the initial identification. Results The target protein was expressed in the supernatant after induction, and the purified protein of interest was initially identified to retain the original biological activity of the native protein. Conclusions The GST / gG-1 fusion protein was obtained, which laid the foundation for the development of a new HSV-1 type specific immunological detection kit with recombinant protein instead of the traditional whole virus as the detection antigen.