摘要
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。
Objective To express HBV PreS2-MBP fusion protein in E. coil then identify and purify the expressed product. Methods Clone the full length of HBV PreS2 gene into prokaryotie expression vector pMAL-P2x by recombinant DNA technique,then transform to E. coil 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 construeted. 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.
出处
《中国生物制品学杂志》
CAS
CSCD
2006年第3期255-258,共4页
Chinese Journal of Biologicals
