乙型脑炎病毒sa1421422株e蛋白主要抗原片段的高效表达.pdfVIP

乙型脑炎病毒 SA14142 株 E 蛋白主要抗原片段的高效表达 1 1 2 1 葛菲菲 ,邱亚峰 ,杨耀武 ,陈溥言 ( 1. 南京农业大学农业部动物疫病诊断与免疫重点开放实验室 ,江苏南京 2 10095 ; 2 . 中国预防医学科学院病毒研究所 ,北京 100052) High Level Expression of Maj or Antigenic Segment of Japanese Encephal itis Virus Strain SA 14142 E Protein GE Feifei1 ,Q IU Yaf en g1 , YAN G Yaowu2 ,C H EN Puyan1 ( 1. Key L aboratory of A nimal D iseases D iag nosis I mm unology of China’s Dep artment of A g ricult ure , N anj ing A g ricult ural Univer ) sity , N anj ing 2 10095 , China; 2. I nstit ute of vi rology . Chinese A cademy of P reventive M edicine. B eij ing , 100052 , China Abstract : Ba sed on a co n st r uct ed p ro karyotic exp re ssion p la smid w hich car rie s a maj or antigenic segment of E gene of J EV st rain SA 14142 in our lab , we exp re ssed t he gene in met hylot rop hic yea st P ich i a p as toris . The segment wa s choo sn ba sed o n an aly si s p rovided by a co mp ut er soft ware becau se of it s higher antigenic index . The seg ment wa s 1113bp , enco din g 37 1 aa . The α gene wa s subcloned into t he vector p P ICZ A . The con st r uct ed p la smid wa s t ran sfor med into yea st X33 by elect rop horation . The reco mbinant t ran sfor mant s were select ed by Zeocin . In duced by t he addition of met ha nol every 24h , t he p ro duct analyzed by SD SPA GE wa s abo ut 44kDa and 50kDa at a yiel d of 290 mg p er lit t er of cult ure . The re sult of We st er n blot ting in dica t ed