A simplified method of constructing infectious clones of begomovirus employing limited restriction enzyme digestion of products of rolling circle amplification
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- 作者：Chia-Ying Wu ; Yi-Chin Lai ; Na-Sheng Lin ; Yau-Heiu Hsu ; Hsin-Tzu Tsai ; Jye-Yann Liao ; Chung-Chi Hu
- 关键词：Begomovirus ; Infectious clone ; Limited restriction enzyme digestion ; Rolling circle amplification
- 刊名：Journal of Virological Methods
- 出版年：2008
- 出版时间：February 2008
- 年：2008
- 卷：147
- 期：2
- 页码：355-359
- 全文大小：546 K
文摘
Most infectious clones of geminiviruses consist of (partial) tandem repeats of viral genomes in the vectors, which usually involve tedious, multi-step assemblies of genomic fragments in the construction process. A simplified procedure was devised to circumvent these problems, which employs limited restriction digestion of multimeric viral genomes produced by rolling circle amplification (RCA), followed by direct cloning into appropriate vectors. The efficiency of the procedure, and infectivity of the dimeric constructs it produced, were demonstrated using three different geminiviruses, namely ageratum yellow vein virus, tomato leaf curl virus, and squash leaf curl virus.