%0 Journal Article
%T The Rescue of H1N1 Subtype Swine Influenza Virus<br>H1N1亚型猪流感病毒的拯救
%A Yaping Peng
%A Hongbo Zhou
%A Chun Li
%A Meilin Jin
%A <br>彭亚平
%A 周红波
%A 李春
%A 金梅林
%J 微生物学报
%D 2008
%I 
%X The swine influenza virus (SIV) strain A/Swine/TianJin/01/2004(H1N1) (A/S/TJ/04) was rescued successfully by an eight-plasmid rescue system. The cDNAs of SIV 8 gene segments were synthesized by RT-PCR and cloned into the RNA polymerase I/II bidirection expression vector PHW2000 independently, resulting in 8 recombinant plasmids. The 8 recombinant plasmids were cotransfected into COS-1 cell, 30 h later TPCK-trypsin was added to 0.5 mg/mL. The COS-1 cell and supernatant were harvested 48 h after cotransfection and were inoculated into the allantoic cavity of 9-day-old specific-pathgen free (SPF) chicken eggs. The allantoic fluid of dead eggs was harvested and passaged 3 generations in SPF chicken eggs to get infective virus. The successful rescue of A/S/TJ/04 SIV was identified by hemagglutination assay, hemagglutination inhibition assay, sequence analysis and electron microscope observation. The successful rescue of SIV built a platform for the research of the relationship between genome structure and function of SIV, the mechanisms of trans-species transmission of influenza virus and for the generation of new SIV as vaccine.
%K swine influenza virus (SIV)
%K bidirection expression vector
%K rescue
%K cotransfection<br>猪流感病毒
%K 双向表达载体
%K 拯救
%K 共转染
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=8BA4BBB1B369FC2CF84A893F404CFF28&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=94C357A881DFC066&sid=E002FF26604EFFAB&eid=0B757E9DCA0EC579&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=0