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| 草鱼(Ctenopharyngodon idellus)呼肠孤病毒(GCRV)VP6相互作用蛋白的筛选 |
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丁燏,李杰,闫秀英,简纪常,吴灶和
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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 广东 湛江 仲恺农业工程学院 广东 广州,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 广东 湛江 仲恺农业工程学院 广东 广州,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 广东 湛江 仲恺农业工程学院 广东 广州,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 广东 湛江 仲恺农业工程学院 广东 广州,仲恺农业工程学院 广东 广州 510000
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| 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pGBKT7以构建其诱饵质粒(pGBKT7-VP6); 再将酵母菌Y2HGold (含pGBKT7-VP6)与酵母菌Y187[含草鱼肾脏细胞(CIK)cDNA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。 |
| 关键词: 酵母双杂交 GCRV096 VP6 蛋白质相互作用 |
| DOI：10.11693/hyhz20140700206 |
| 分类号:Q939.94；S942.2 |
| 基金项目: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 |
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| SCREENING OF THE PROTEINS INTERACTED WITH VP6 IN GRASS CARP (CTENOPHARYNGODON IDELLUS) REOVIRUS |
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DING Yu,LI Jie,YAN Xiu-Ying,JIAN Ji-Chang and WU Zao-He
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Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Fisheries College,Guangdong Ocean University,Zhanjiang,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Fisheries College,Guangdong Ocean University,Zhanjiang,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Fisheries College,Guangdong Ocean University,Zhanjiang,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Fisheries College,Guangdong Ocean University,Zhanjiang,Zhongkai University of Agriculture and Engineering, Guangzhou, Guangdong 510000,China
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| Abstract: |
| 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 pGBKT7 to construct pGBKT7-VP6. The yeast Y2HGold containing the recombinant vector of VP6 gene was mated with the yeast Y187 pre-transformed with cDNA 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 cDNA sequences were identified. The protein encoded by a cDNA fragment has higher homology with glyceraldehyde-3-phosphate dehydrogenase. The enzyme may be important for GCRV096 to entry into the cells of Ctenopharyngodon idellus. |
| Key words: yeast two-hybrid system GCRV096 VP6 protein interaction |