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Generation of stable infectious clones of plant viruses by using Rhizobium radiobacter for both cloning and inoculation

文献类型： 外文期刊

作者： Tuo, Decal 1 ; Fu, Lanlan 1 ; Shen, Wentao 1 ; Li, Xiaoying 1 ; Zhou, Peng 1 ; Yan, Pu 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Key Lab Biol & Genet Resources Trop Crops, Minist Agr, Haikou 571101, Hainan, Peoples R China

关键词： Plant viruses;Infectious clone;Rhizobium radiobacter;PRSV;PLDMV;E. coif;Instability

期刊名称：VIROLOGY （ 影响因子：3.616； 五年影响因子：3.967 ）

ISSN： 0042-6822

年卷期： 2017 年 510 卷

页码：

收录情况： SCI

摘要： A novel Rhizobium radiobacter (synonym Agrobacterium tumefaciens)-mediated approach was developed to generate stable infectious clones of plant viruses. This method uses R. radiobacter for both cloning and inoculation of infectious clones, bypassing the requirement of cloning in E. coli to avoid the instability. Only three steps are included in this method: (i) construct viral genome-encoding plasmids in vitro by one-step Gibson assembly; (ii) transform the assembled DNA products into R. radiobacter; (iii) inoculate plants with the R. radiobacter clones containing the viral genome. Stable infectious clones were obtained from two potyviruses papaya ringspot virus (PRSV) and papaya leaf distortion mosaic virus (PLDMV) using this method, whereas attempts utilizing "classical" E. coil cloning system failed repeatedly. This method is simple and efficient, and is promising for a wide application in generation of infectious clones of plant virus, especially for those which are instable in E. colt.

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