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赵云

作品数:2 被引量:5H指数:1
供职机构:中国疾病预防控制中心病毒病预防控制所更多>>
发文基金:卫生部科学研究基金国家自然科学基金更多>>
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新疆出血热病毒糖蛋白基因的克隆与表达被引量:1
2002年
On the basis of sequencing and analyzing of the whole M genes(encoding viral membrane antigen glycoprotein) of three Crimean Congo hemorrhagic fever viruses(CCHFV),the Chinese isolates(Xinjiang hemorrhagic fever virus,XHFV),we first expressed the glycoprotein (GP) gene of the prototype human origin XHFV strain BA66019 in eukaryotic cells and investigated the expression profiles.Three eukaryotic expression plasmids were constructed starting from ATG 78 (the first start codon of the deduced entire XHFv GP precursor gene positioned between the 78 th ~80 th nucleotides),ATG 93 and ATG 3084 (the potential start codon for G1 precursor gene).The constructs were transfected to COS-7 cells and the expressed products were characterized as membrane-bound proteins which could induce cell fusion This was much more apparent for recombinant plasmid with ATG 3084 A recombinant baculovirus was further created harboring full length GP gene(starting from ATG 78 ) and the expression could also result in the membrane fusion as well as swelling of the infected Sf9 cells The insect cell expressed G1 was smaller in M W than natural G1 (approximately 67kD) on SDS-PAGE and no recombinant G2 band was detectable Western-blot only detected the native G1,while there was no specific corresponding band of recombinant G1 These data suggested that G1 was structurally and functionally important in XHFV and the glycosylation may have great influence on M W as well as antigenicity This study provides a foundation for the future study of viral pathogenesis ,antigenicity and immunity,that are the theoretical and experimental backgrounds for vaccine
马本江杭长寿赵云王世文解燕乡
关键词:新疆出血热病毒糖蛋白基因克隆
以新型杆状病毒载体在昆虫、哺乳动物细胞中表达克里米亚-刚果出血热病毒核蛋白基因被引量:4
2002年
目的 构建可以在哺乳动物细胞中高效表达的新型杆状病毒载体 ,并利用其将克里米亚 刚果出血热病毒 (CCHFV)中国分离株 (新疆出血热病毒 ,XHFV)BA8816 6的核蛋白 (NP)基因在昆虫和哺乳动物细胞中进行表达。方法 将人巨细胞病毒 (CMV)立即早期 (IE)启动子连接至杆状病毒载体pFastBac1多角体启动子下游形成新载体pCB1,然后将XHFVNP基因克隆至该载体 ,通过重组质粒转染和病毒感染 ,检测其在哺乳动物细胞 (COS 7和Vero)及昆虫细胞中的表达。结果 连接至pCB1的XHFVNP基因均能在相应的细胞中获得良好表达 ;以重组杆状病毒感染的Vero细胞可以作为抗原检测XHF血清 ,与ELISA的检测结果完全一致 ,并与临床诊断有很好的平行性。结论 新型杆状病毒载体能够驱动外源基因在昆虫和哺乳动物细胞中高效表达 ,不仅能方便快速地制备诊断抗原 ,还具有发展重组病毒疫苗和基因治疗的潜力。
马本江杭长寿赵云王世文解燕乡
关键词:核蛋白基因基因表达昆虫哺乳动物
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