重组H9N2禽流感病毒的构建及其在不同细胞中的复制特性

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (4): 480-484.

重组H9N2禽流感病毒的构建及其在不同细胞中的复制特性

贾倩楠¹,赵丽丽²,邓涛²,曹萌萌²

1. 中国医学科学院/北京协和医学院病原生物学研究所
2. 中国医学科学院病原生物学研究所

收稿日期:2014-12-19 修回日期:2015-01-13 出版日期:2015-04-05 发布日期:2015-04-08
通讯作者: 曹萌萌 E-mail:caommeng@ipbcams.ac.cn
基金资助:
病毒性疾病感染组学关键技术平台十二五重大专项

Generation of an H9N2 avian influenza virus and characterization of its replication properties in different cells

Received:2014-12-19 Revised:2015-01-13 Online:2015-04-05 Published:2015-04-08

摘要/Abstract

摘要： 目的利用反向遗传学技术构建具有感染性的H9N2亚型禽流感病毒，并揭示其在不同细胞中的复制特性。方法以A/Quail/HongKong/G1/97(H9N2)禽流感病毒基因组RNA为模板，用RT-PCR技术获得该病毒的8条完整基因片段，并分别将它们插入到pHW2000载体上，最终在293T细胞中包装产生重组H9N2病毒。并将重组的H9N2病毒感染不同细胞，观察病毒在不同细胞上的增殖状况来揭示该病毒在不同细胞上的复制特性。结果应用流感病毒8质粒反向遗传学技术，成功获得重组病毒，并揭示了该毒株在不同细胞(A549, MDCK, Vero)中的复制特性,发现A549人肺腺癌细胞更适宜H9N2病毒的正常复制。此外还分析总结了该毒株的与病毒毒力相关的重要位点特征。结论成功建立了H9N2禽流感病毒的反向遗传系统，该系统的建立为在分子水平研究H9N2病毒以及制备H9N2禽流感疫苗奠定了基础。

关键词: 禽流感病毒, 反向遗传学, 复制特性

Abstract: Objective Generation of an H9N2 avian influenza virus with reverse genetics and characterization of its replication properties in different cells. Methods Eight genes of avian influenza virus A/Quail/HongKong/G1/97(H9N2) were amplified by RT-PCR technique and inserted into the pHW2000 vectors. The recombinant virus was rescued by cell transfection and the virus replication properties were analyzed in A549, MDCK and Vero cells. Results Using eight plasmid-based reverse genetics, the recombinant Avian H9N2 influenza virus was rescued. It is observed that A549 cells were more suitable for the H9N2 virus replication than MDCK cells and Vero cells. The molecular virulence markers of the virus were also summarized with bioinformatics technology. Conclusion The eight plasmid-based reverse genetic system is successfully set up for the avian H9N2 virus. The system could not only facilitate genetic manipulation of the H9N2 virus but also provide a platform in studying the virus and developing avian influenza vaccine.

Key words: Avian influenza virus, Reverse genetics, Growth property

中图分类号:

R373.1+3

贾倩楠赵丽丽邓涛曹萌萌. 重组H9N2禽流感病毒的构建及其在不同细胞中的复制特性[J]. 基础医学与临床, 2015, 35(4): 480-484.