作　　者： ; ; (闫秀英）; (简纪常）; (吴灶和）;

机构地区： 广东海洋大学水产学院

出　　处： 《海洋与湖沼》 2014年第6期1295-1299,共5页

摘　　要： 为筛选与草鱼呼肠孤病毒GCRV096 VP6发生相互作用的宿主蛋白,先将VP6基因的PCR扩增产物,克隆至酵母表达载体p GBKT7以构建其诱饵质粒(p GBKT7-VP6);再将酵母菌Y2HGold(含p GBKT7-VP6)与酵母菌Y187[含草鱼肾脏细胞(CIK)c DNA文库质粒]进行人工诱导融合,然后经选择培养筛选阳性克隆。结果共筛选到4株阳性克隆,经测序、生物信息学分析,分别属于两条不同的核苷酸序列,其中之一所编码的产物与GAPDH(3-磷酸甘油醛脱氢酶)同源性较高。可见,该酶极可能在GCRV096侵染宿主的初期发挥着重要作用。 To screen and identify the proteins that interact with VP6 of GCRV096 strain, VP6 gene was amplified by polymerase chain reaction(PCR) and then inserted into bait plasmid p GBKT7 to construct p GBKT7-VP6. The yeast Y2 HGold containing the recombinant vector of VP6 gene was mated with the yeast Y187 pre-transformed with c DNA library plasmid of CIK. Four positive clones were obtained by screening the diploid yeast cells. The inserted fragments were sequenced and analyzed by bioinformatic method. The results show that two different c DNA sequences were identified. The protein encoded by a c DNA fragment has higher homology with glyceraldehyde-3-phosphate dehydrogenase. The enzyme may be important for GCRV096 to entry into the cells of Ctenopharyngodon idellus.

关 键 词： 酵母双杂交 蛋白质相互作用

领　　域： [农业科学] [农业科学]