摘要
应用反向遗传技术将含有1998年中国大陆分离株H9N2亚型禽流感病毒(Avianinfluenzavirus,AIV)的8个基因片段的质粒共转染COS_1细胞,产生了与野生病毒生物学特性相同的H9N2亚型AIV。将A Chicken Shanghai F 98(CK SH F 98)株H9N2亚型AIV的8个基因组cDNA分别克隆到polⅠ_polⅡ转录 表达载体pHW2 0 0 0中,构建成8个转录表达载体重组质粒。将这8个质粒共转染COS_1细胞,2 4h后收获细胞及上清接种SPF鸡胚,4 8h后收取鸡胚尿囊液继续进行鸡胚传代,产生能致死鸡胚的病毒。经血凝、血凝抑制试验、序列分析和电镜观察,证实产生了CK SH F 98(H9N2 )株AIV。
Eight-plasmid system was used for the generation of Avian influenza virus(AIV) strain A/Chicken/Shanghai/F/98 (H9N2) which was isolated in China in 1998. In this plasmid_based expression system, viral cDNA was inserted beteen the RNA polymeraseⅠ(polⅠ) promoter and terminator sequences. The entire polⅠtranscription unit was flanked by an RNA polymerase Ⅱ(polⅡ) promoter and a poly (A) site. Twenty_four hours after the transfection of eight expression plasmid into cos1 cells, the supernatant and cos1 cells transfected were inoculated into the allantoic cavity of 10_day_old specific_pothgen_free(SPF) chicken eggs . The HA titer was determined after passage of the rescued virus in chicken eggs, and as high as that of the parental wild_type virus.
出处
《微生物学报》
CAS
CSCD
北大核心
2005年第3期373-376,共4页
Acta Microbiologica Sinica
基金
江苏省"十五"科技攻关项目 (BE2 0 0 2 0 16)
农业部资助项目 (农办牧 2 0 0 183 )~~
