新疆出血热病毒S基因在真核系统的克隆和表达  

Cloning and expression of XHFV S gene in eukaryote expression system

作　　者：唐青[1] 于凤刚[1] 赵秀芹[1] 陶晓霞[1] Anna Papa A Antoniadis 杭长寿[3] 

机构地区：[1]中国预防医学科学院流行病学微生物学研究所 [2]希腊亚里士多德大学医学院 [3]中国预防医学科学院病毒所

出　　处：《中华微生物学和免疫学杂志》2002年第1期28-32,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 ( 39870 6 80 )

摘　　要：目的　在真核系统表达XHFVS基因片段 ,制备安全、特异、便于操作的S蛋白。方法设计引物 ,扩增得到S基因编码片段 ,用杆状病毒表达载体pFastBacHTb克隆S基因 ,并在昆虫细胞中表达S基因。结果　地高辛探针杂交和PCR扩增结果表明成功地构建了含XHFVS基因片段的pBac Sxhf质粒。SDS PAGE分析表达产物在相对分子质量 (Mr)为 6 6× 10 3 下方出现 1条明显的蛋白条带 ,Westernblot发现该蛋白带可以与抗XHF病毒核蛋白单克隆抗体发生反应。结论　构建的含XHFVS基因的重组杆状病毒可以在昆虫细胞表达S蛋白 。Objectives Expressing of Xinjiang hemorrhagic fever virus(XHFV) S gene in an eukaryote expression system and getting a safe, specific and easy manipulating antigen. Methods To design primers for amplifying XHFV S coding segment and get S coding segment by PCR, clone this segment by baculovirus expression vector with 6x histiding-affinity tag and then use a recombinant plasmid (pBac-Sxhf) to transfect Sf9 cells for the expression of S gene. Result The presence of S gene of XHFV in pBac-Sxhf recombinant plasmid was proven by hybridization of Digoxin probe and PCR. Expressed S protein was analyzed by SDS-PAGE and the Ag-Ab reactivity between the expressed S protein and McAb against XHFV was confirmed by Western blot. Conclusion Recombinant baculoviruses secrete XHFV S protein and are specific reaction with McAb against S protein of XHFV.

关 键 词：新疆出血热 杆状病毒 S基因 重组 质粒 表达蛋白 克隆 

分 类 号：R346[医药卫生—基础医学]