摘要
目的 :制备和鉴定汉滩病毒 (HTNV)S基因 重组痘苗病毒 ,为进一步建立肾综合征出血热 (HFRS)患者HTNV核衣壳蛋白特异性细胞毒T淋巴细胞 (CTL)的靶细胞奠定基础。 方法 :用野生型痘苗病毒和HTNVS基因 重组痘苗病毒分别感染Vero细胞 ,进行病毒扩增和滴度测定。用PCR和斑点杂交鉴定重组痘苗病毒基因组中的HTNVS基因 ,用间接免疫荧光和westernblot检测核衣壳蛋白的表达。 结果 :制备了含HTNVS基因的重组痘苗病毒 ,滴度为 4 .0 2× 10 7。该重组痘苗病毒基因组中存在HTNVS基因 ,它在Vero细胞中能有效地表达核衣壳蛋白。 结论 :制备了较高滴度、能有效表达HTNV核衣壳蛋白的重组痘苗病毒。
Objective:Preparing and identifing recombinant vaccinia virus with Hantaan virus S gene, to lay the foundation for the establishment of specific cytotoxic T lymphocytes (CTL) target cells expressing nucleocapsid protein of Hantaan virus. Methods: Vero cells were infected by wild vaccinia virus and recombinant vaccinia virus with Hantaan virus S gene, respectively. Viruses were multiplied and tittered. Hantaan virus S gene in recombinant vaccinia virus genome was identified using PCR and dot hybridization. Expression of nucleocapsid protein was detected using indirect immunofluorescent assay (IFA) and Western blot. Resusts:Recombinant vaccinia viruses with Hantaan virus S gene have been prepared. Virus titer is 4.02×10 7 . It can effectively express nucleocapsid protein in Vero cells. Conclusion:Recombinant vaccinia viruses with higher titer were prepared successfully and it can effectively expressed nucleocapsid protein of Hantaan virus.
出处
《医学研究生学报》
CAS
2004年第1期18-20,24,共4页
Journal of Medical Postgraduates
