摘要
为研究WSSV的粘附蛋白VP37相关蛋白的基因,应用酵母双杂交系统Ⅲ筛选中国明对虾(Fenneropenaeus chinensis)鳃细胞全长cDNA文库。构建VP37 基因的重组载体PGBKT7 VP37,把它转化酵母Y187后与含文库质粒的酵母AH109配合,在营养缺陷培养基上进行双杂交筛选。经过PCR鉴定确定了1个阳性克隆。测定该段cDNA序列并进行生物信息学分析,结果显示,此cD NA片段编码的蛋白与蛋白酶的同源性较高,该蛋白在WSSV识别并结合宿主的过程中所起的作用还有待于进一步研究。成功筛选了VP37相关蛋白的cDNA片段,为研究WSSV入侵以及感染致病的机制等方面提供了新线索。
To research the genes which encode the virus attachment protein 37 of WSSV (White Spot Syndrome Virus), the yeast two-hybrid system ш was used to screen the full-length cDNA library from gills of Fenneropenaeus chinensis. The recombinant vector of gene VP37 was constructed and transformed into yeast Y187. It was mating with AH109 that pre-transformed with cDNA library plasmids, and the diploid yeast cells were screened on dropout medium. After the identification of PCR, one positive clone was obtained. This cDNA fragment was sequenced and assayed in bioinformatics. The results showed that the proteins encoded by the cDNA fragment have higher homology with protease, The function of the protein in the process of the conjunction of WSSV and its host need to be further researched. The cDNA fragmentinterrelating with the gene of VP37 is obtained successfully and the results provided some new clues for studying the mechanism of the intrusion and infection of WSSV.
出处
《海洋水产研究》
CSCD
北大核心
2005年第2期73-78,共6页
Marine Fisheries Research
基金
国家重点基础研究发展规划项目(G1999012002)
国家自然科学基金项目(30271020)
青岛市自然科学基金项目(03 2 JZP 10)
山东省自然科学基金项目(Q2002D02)
中国水产科学研究院科研基金项目(2003-青-6)共同资助
