黄瓜绿斑驳花叶病毒全长及绿色荧光蛋白标记的cDNA克隆的农杆菌接种系统的开发
Development of an agroinoculation system for full-length and GFP-tagged cDNA clones of cucumber green mottle mosaic virus.
作者信息
Zheng Hongying, Xiao Caili, Han Kelei, Peng Jiejun, Lin Lin, Lu Yuwen, Xie Li, Wu Xiaohua, Xu Pei, Li Guojing, Chen Jianping, Yan Fei
机构信息
State Key laboratory Breeding Base for Sustainable Control of Plant Pest and Disease, Key Laboratory of Biotechnology in Plant Protection of Ministry of China and Zhejiang Province, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China.
College of Chemistry and Life Science, Zhejiang Normal University, Jinhua, 321004, China.
出版信息
Arch Virol. 2015 Nov;160(11):2867-72. doi: 10.1007/s00705-015-2584-y. Epub 2015 Sep 1.
The complete 6243-nucleotide sequence of a cucumber green mottle mosaic virus (CGMMV) isolate from bottle gourd in Zhejiang province, China, was determined. A full-length cDNA clone of this isolate was constructed by inserting the cDNA between the 35S promoter and the ribozyme in the binary plasmid pCB301-CH. A suspension of an Agrobacterium tumefaciens EHA105 clone carrying this construct was highly infectious in Nicotiana benthamiana and bottle gourd. Another infectious clone containing the green fluorescence protein (GFP) reporter gene was also successfully constructed. This study is the first report of the efficient use of agroinoculation for generating CGMMV infections.
测定了来自中国浙江省瓠瓜的一株黄瓜绿斑驳花叶病毒（CGMMV）分离物的完整6243个核苷酸序列。通过将该cDNA插入二元质粒pCB301-CH的35S启动子和核酶之间，构建了该分离物的全长cDNA克隆。携带此构建体的根癌农杆菌EHA105克隆悬浮液在本氏烟草和瓠瓜中具有高度传染性。还成功构建了另一个含有绿色荧光蛋白（GFP）报告基因的感染性克隆。本研究首次报道了利用农杆菌接种高效产生CGMMV感染。
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