重组H8N4亚型禽流感病毒的拯救

THE RESCUE OF RECOMBINANT H8N4 AVAIN INFLUENZA VIRUS

将H8N4亚型流感病毒的HA和NA基因插入双向转录/表达载体pHW2000中,并与本实验室保留的适应毒株A/PR/8/34(H1N1)的6个质粒pHW2000-PB2、pHW2000-PB1、pHW2000-PA、pHW2000-NP、pHW2000-M、pHW2000-NS共转染293T和MDCK混养细胞,在细胞内部完成病毒粒子的组装,然后接种SPF鸡胚,经3代扩毒后,成功拯救了H8N4亚型重组流感病毒,并对其进行鉴定及生物学特性分析。本研究结果为该亚型禽流感病毒变异机理研究及疫苗的研制等奠定了基础。

In the present study, HA and NA genes of Avian influenza virus （AIV） H8N4 were inserted to the transcription/expression plasmid pHW2000, and then co-transfected the mixture of 293T and MDCK cultures with the following six plasmids of the adaptive strain ofA/PR/8/34（H1N1）retainedinourlaboratory： pHW2000-PB2, pHW2000-PB1, pHW2000-PA、 pHW2000-NP, pHW2000-M and pHW2000-NS. After completion of assembly, viruses were inoculated into chick embryos and passaged three times. The resulting custom-crafted HSN4 subtype virus was obtained and characterized. The present study provided useful information for future studies on AIV variation and vaccine research.