| 摘要: |
| 采用酵母双杂交技术, 对草鱼呼肠孤病毒096(GCRV096)非结构蛋白NS38相互作用的蛋白进行了筛选, 并对阳性克隆中的插入序列进行测序与生物信息学分析。结果表明, 含有重组质粒pGBKT7-NS38的酵母菌Y2HGold与含有草鱼肾脏细胞(CIK)cDNA文库质粒的酵母菌Y187融合成功, 并筛选得到3株阳性克隆; 发现两条不同的插入序列, 其中一条序列编码的蛋白与40S核糖体蛋白S16亚基的同源性高达99%, 而另一序列相应的蛋白不存在匹配的蛋白。该结果为进一步深入研究NS38蛋白在GCRV096复制过程中的生物学功能奠定了基础。 |
| 关键词: GCRV096 NS38 蛋白质相互作用 酵母双杂交 |
| DOI：10.11693/hyhz201302006006 |
| 分类号: |
| 基金项目:国家973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 |
附件 |
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| SCREENING THE PROTEINS INTERACTED WITH NS38 IN GRASS CARP REOVIRUS BY YEAST TWO-HYBRID SYSTEM |
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LI Jie1, YAN Xiu-Ying1, DING Yu1, WU Zao-He2, JIAN Ji-Chang1, XIE Ji-Guo1
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1.Guangdong Provincial Key Laboratory ofPathogenic Biology and Epidemiology for Aquatic Economic Animals, Fisheries College, Guangdong Ocean University;2.Zhongkai University of Agriculture and Engineering
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| Abstract: |
| The proteins interactive with non-structural protein NS38 of GCRV096 (grass carp reovirus) was screened in a yeast two-hybrid system, and the inserted fragments in positive clones were sequenced and analyzed bioinformatically. The results show that yeast Y2HGold containing the recombinant vector of NS38 gene was successfully fused with yeast Y187 that pre-transformed with cDNA library plasmid of CIK. Three positive clones were obtained, and two inserted cDNA sequences were identified. The protein encoded by a cDNA fragment has higher homology of 99% with S16 subunit of 40S ribosomal protein. These results may provide a basis for further study on the biological function of NS38 in the duplication of GCRV096. |
| Key words: GCRV096 NS38 protein interaction yeast two-hybrid system |
