HBV PreS2-MBP融合蛋白质粒构建及在大肠杆菌中的表达

目的在大肠杆菌pMALP2x系统中表达HBVPreS2MBP融合蛋白,并对其进行鉴定和纯化。方法利用DNA重组技术,将全长的HBVPreS2蛋白基因克隆至原核表达载体pMALP2x中,转化大肠杆菌DH5α,IPTG诱导表达。表达产物经SDSPAGE和Westernblot检测,并经阴离子交换层析、AmyloseResin亲和层析纯化。结果成功构建了重组载体pMALP2x/S2,诱导蛋白为MBP标记的可溶性的PreS2MBP融合蛋白,具有良好的抗原性,可经AmyloseResin亲和层析纯化。结论pMALP2x系统可成功表达PreS2MBP融合蛋白。

Construction of Recombinant Plasmid for Expression of HBV PreS2-MBP Fusion Protein in E. coli

Objective To express HBV PreS2-MBP fusion protein in E.coli then identify and purify the expressed product.Methods Clone the full length of HBV PreS2 gene into prokaryotic expression vector pMAL-P2x by recombinant DNA technique,then transform to E.coli for expression under induction of IPTG.Identify the expressed product by SDS-PAGE and Western blot then purify by anion-exchange and Amylose Resin affinity chromatography.Results Recombinant plasmid pMAL-P2x/S2 was successfully constructed.The MBP-labeled PreS2-MBP fusion protein with good antigenicity was expressed in a soluble form and successfully purified by Amylose Resion affinity chromatography.Conclusion HBV PreS2-MBP fusion protein was successfully expressed in pMAL-P2x system.