戊型肝炎病毒结构区基因在大肠杆菌中的表达

目的获得可用于研制戊型肝炎疫苗的基因工程抗原。方法用非融合蛋白表达载体ｐＥＴ１１ａ，表达戊型肝炎病毒（ＨＥＶ）第二读码框区（ＯＲＦ２）２２４６６０ａａ段蛋白。结果得到分子量为５００００的表达产物，经Ｗｅｓｔｅｒｎｂｌｏｔ证实，表达产物具有ＨＥＶ特异抗原性。结论该基因工程抗原可能具有制备戊型肝炎疫苗的前景。

Expression of hepatitis E virus structural gene in E. coli.

Objective To obtain recombinant antigen for development of vaccine against hepatitis E virus. Methods Amplified the structural gene (5 8167 126 nt) by PCR. The upstream primer was 5CCATATGAATTCAATAACCTC3 and the downstream primer was 5GGGATCCTATAACTCCCGAGT3. Cut the PCR product with Nde I and BamHI, then inserted this fragment into the plasmid pET11 where a cut was by the same restriction endonucleases. The expression plasmid named pEa47 was transformed into E. Coli BL21. The recombinant strains were grown at 37 and induced by IPTG. The recombinant protein was confirmed by Western blot analysis using serum from hepatitis E patient. Results The structural gene of hepatitis E virus from open reading frame 2,224660 aa, was expressed in E. Coli BL21. Western blot assay showed that the expressed 50 000 recombinant protein specifically reacted with the serum antibody from the hepatitis E patient. Conclusion The protein might be useful to develop vaccine against hepatitis E virus infection.