National Influenza Centre and Department of Virology, Erasmus Medical Centre, Rotterdam, The Netherlands.
Vaccine. 2010 Aug 16;28(36):5803-9. doi: 10.1016/j.vaccine.2010.06.072. Epub 2010 Jul 13.
Reverse genetics can be used to produce recombinant influenza A viruses containing virtually every desired combination of hemagglutinin (HA) and neuraminidase (NA) genes using the virus backbone of choice. Here, a repository of plasmids and recombinant viruses representing all contemporary Eurasian HA and NA subtypes, H1-H16 and N1-N9, was established. HA and NA genes were selected based on sequence analyses of influenza virus genes available from public databases. Prototype Eurasian HA and NA genes were cloned in bidirectional reverse genetics plasmids. Recombinant viruses based on the virus backbone of A/PR/8/34, and containing a variety of HA and NA genes were produced in 293T cells. Virus stocks were produced in MDCK cells and embryonated chicken eggs. These plasmids and viruses may be useful for numerous purposes, including influenza virus research projects, vaccination studies, and to serve as reference reagents in diagnostic settings.
反向遗传学可用于使用所选病毒骨架，产生包含几乎所有所需血凝素 (HA) 和神经氨酸酶 (NA) 基因组合的重组甲型流感病毒。在这里，建立了一个包含所有当代欧亚 HA 和 NA 亚型（H1-H16 和 N1-N9）的质粒和重组病毒库。根据公共数据库中提供的流感病毒基因的序列分析选择了 HA 和 NA 基因。将原型欧亚 HA 和 NA 基因克隆到双向反向遗传学质粒中。在 293T 细胞中基于 A/PR/8/34 的病毒骨架产生了含有多种 HA 和 NA 基因的重组病毒。在 MDCK 细胞和鸡胚中生产病毒储备。这些质粒和病毒可能有多种用途，包括流感病毒研究项目、疫苗研究以及作为诊断环境中的参考试剂。
