文献类型：期刊文献

中文题名：草鱼(Ctenopharyngodon idellus)呼肠孤病毒(GCRV)VP6相互作用蛋白的筛选

英文题名：SCREENING OF THE PROTEINS INTERACTED WITH VP6 IN GRASS CARP(CTENOPHARYNGODON IDELLUS) REOVIRUS

作者：丁燏[1];李杰[1];闫秀英[1];简纪常[1];吴灶和[2]

机构：[1]广东省水产经济动物病原生物学及流行病学重点实验室 广东海洋大学水产学院,湛江524025;[2]仲恺农业工程学院,广州510000

年份：2014

卷号：45

期号：6

起止页码：1295

中文期刊名：海洋与湖沼

外文期刊名：Oceanologia Et Limnologia Sinica

收录：CSTPCD、、北大核心2011、Scopus、北大核心、CSCD、CSCD2013_2014

基金：国家重点基础研究发展计划"水生动物呼肠孤病毒的致病机理与保护性抗原研究";2009CB118704号

语种：中文

中文关键词：酵母双杂交;GCRV096;VP6;蛋白质相互作用

外文关键词：yeast two-hybrid system;;GCRV096;;VP6;;protein interaction

中文摘要：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。

外文摘要：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.