Several H11N2 subtype of Avian influenza A viruses were isolated from aquatic birds in live bird markets when we surveyed the ecology of the influenza in East China for more than two years and identified by specific *...
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Several H11N2 subtype of Avian influenza A viruses were isolated from aquatic birds in live bird markets when we surveyed the ecology of the influenza in East China for more than two years and identified by specific *** hemagglutinin(HA) and neuraminidase(NA) genes of one representative virus named A/Duck/Yangzhou/44/2002(H11N2)(DYZ/44/02) was *** results showed the HA nucleotide sequence of DYZ/44/02 has high identity with Dk/England/56(H11N6) and the NA nucleotide sequence of DYZ/44/02 has more than 95% sequence homology with Dk/Hokkaido/49/98(H9N2).Sequencing and phylogenetic analysis of the N2 NA genes of DYZ/44/02 revealed that the NA gene of DYZ/44/02 has close relationships with that of Ck/Korea/MS96/96-like H9N2 virus and are distinct from those of Ck/Beijing/94(H9N2).The sequence of cleavage site of DYZ/44/02 consists of a single arginine,as is the case with most other hemagglutinins exhibiting low susceptibility to proteolytic activation.
运用PCR技术,分别从大肠杆菌TB1、107/86和C83907菌株中扩增出不含信号肽序列的Stx 2e B亚单位基因(stx 2e B)、F18茵毛的A亚单位基因(fedA)和F4茵毛亚单位基因(faeG),将这3个基因的PCR扩增产物按预定阅读框架克隆入pGEX-6P-1载体的谷...
详细信息
运用PCR技术,分别从大肠杆菌TB1、107/86和C83907菌株中扩增出不含信号肽序列的Stx 2e B亚单位基因(stx 2e B)、F18茵毛的A亚单位基因(fedA)和F4茵毛亚单位基因(faeG),将这3个基因的PCR扩增产物按预定阅读框架克隆入pGEX-6P-1载体的谷胱甘肽S-转移酶(GST)基因的下游,对构建的重组质粒pFSFaeG进行酶切分析和核苷酸序列分析,结果表明,插入的片段与预期一致,且阅读框正确。重组质粒在宿主茵BL21中的表达产物经SDS-PAGE初步分析,发现表达产物与谷胱甘肽转移酶相融合,重组融合蛋白的分子量约为80kD。用表达FedA-Stx 2e B-FaeG的重组茵BL21(pFSFaeG)灭活苗对5周龄的ICR小鼠进行腹腔免疫,结果显示经重组菌免疫后的小鼠血清中,产生了分别针对F4、F18、Stx2e抗原特异性的IgG,相应的抗体水平从免疫后7d开始逐步升高,在第二次免疫后21d左右达到了较高的水平,并且在攻毒后对小鼠提供了较好的保护,免疫保护率均达80%以上。
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