期刊论文

中文

中国人兽共患病学报

1002-2694

2007

23

9

899-902,906

10.3969/j.issn.1002-2694.2007.09.013

国家科技支持计划资助项目（2006BAD06A01）；

本校为第一机构

目的 构建H5N1亚型禽流感病毒NP基因的酵母双杂交诱饵载体,验证其在酵母中的表达并检测其自激活作用.方法以PCR法从pGEMT/H5NP扩增NP基因编码区序列,将其定向克隆到PGBKT7载体,经测序鉴定后,PEG/LiAc法转化酵母菌株AH109,用表型筛选法及颜色筛选法检测其自激活作用同时Western blot验证诱饵蛋白的表达.结果获得NP编码区基因,并成功构建酵母双杂交系统中的诱饵载体pGBKT7-NP,对宿主细胞酵母菌株AH109无毒性和自激活作用,并能在酵母细胞中稳定表达.结论诱饵载体pGBKT7-NP可用于GAL4酵母双杂交系统钓取与禽流感病毒核蛋白相互作用的蛋白.

To express of nucleoprotein（NP） of avian influenza virus （H5N1） in yeast two-hybrid system and to detect its self-activation in this system, the complete encoding gene of NP was amplified from a plasmid pGEMT/H5NP by PCR and it was inserted into pGBKT7 vector according to the reading frame. The bait vectors were transformed into the yeast strain AH109 by PEG/LiAc method and its self-activation was tested by both the phenotype assay and the color assay,then expression of Ga14-NP fusion proteins identified by Western blot. In this way, the ba...