摘要
目的用基因工程的方法获得乙型肝炎病毒核心抗原(HBcAg)编码区的基因片段,构建重组质粒并在大肠杆菌中表达抗原蛋白。方法用PCR法扩增HBcAg基因片段,构建含有HBcAg基因的克隆质粒及表达质粒,在大肠杆菌BL21(DE3)plys中以IPTG诱导重组蛋白的表达,并用Western blot对重组蛋白进行检测。结果重组HBcAg在大肠杆菌中得到表达,Western blot检测显示在预期位置出现特异性条带。结论成功构建HBcAg原核表达系统并获得重组HBcAg,为该重组蛋白的相关功能研究奠定了基础。
Objective To obtain the gene fragment encoding HBcAg,construct recombinant plasmid and express the recombinant HBcAg protein in E.coli using genetic engineering methods.Methods The HBcAg gene fragment was amplified by PCR and the cloning and expression plasmids including HBcAg gene were constructed.Then the recombinant plasmid was transformed into E.coli BL21（DE3）host cell.The HBcAg fusion protein was expressed under the induction of IPTG and identified by Western blot.Results Recombinant HBcAg was expressed in E.coli.The special band of recombinant protein was showed at expectant position by Western blot.Conclusion A prokaryotic expression system for HBcAg gene is successfully constructed and the recombinant HBcAg protein is obtained.It lays a foundation of further study on the function of the recombinant protein.
出处
《检验医学与临床》
CAS
2010年第17期1802-1803,共2页
Laboratory Medicine and Clinic
