禽流感病毒M1蛋白的表达及其免疫反应性分析.pdfVIP

禽流感病毒 M1 蛋白的表达及其免疫反应性分析 宋建领 ,王金萍 ,胡媛媛 ,张富强 ( 云南省热带亚热带动物病毒病重点实验室 ,云南昆明 650224) Expression of M1 protein of Avian Infl uenza Virus and Analysis of Its Immunoreactivity SON G J ianling , WAN G J unping , HU Yuanyuan , ZHAN G Fuqian g ( ) Yunnan T rop ical an d S ubt rop ical A ni m al Vi rus D iseases L aboratory , K unm i ng 650224 , Chi na Abstract : A p air of p rimer s were de signed ba sed on M 1 gene sequence of know n H5N 1 Avian in fluenza vir u s. M 1 gene wa s cloned f ro m tot al RN A , ext ract ed f ro m ti ssue of H5N 1 subt yp e vir u s inoculat ed embryo by rever se t ran scrip t a se polymera se chain reaction u sin g high p roof readin g polymera se ( Pyobe st TM DN A Polymera se) , and exp re ssed u sin g Invit ro gen champ ion TM p E T di rectional TO PO exp re ssion sy st em . Reco mbinant p rot ein cont ainin g polyhi stidine (6x Hi s) t ag in Nt er minal abo ut 29 . 8kDa in size ,wac obt ained and p urified . It s immunoreactivit y wa s analyzed by We st er n blot , EL ISA and blockin g EL ISA u sing MA b and po sitive ser um . The re sult s showed reco mbinant M 1 p rot ein can bind to MA b an d po sitive ser um wit h sp ecificit y , and t hi s binding reactivit y can be blocked by nat ural viral antigen . Thi s indicat e s t hat t he reco mbinant M 1 p rot ein po sse sse s goo d immunoreactivit y . Key words : Avian influenza viru s ; M 1 p