A simplified method of constructing infectious clones of begomovirus employing limited restriction enzyme digestion of products of rolling circle amplification

被引：28

作者：

Wu, Chia-Ying ^{[1
]}

Lai, Yi-Chin ^{[1
]}

Lin, Na-Sheng ^{[2
]}

Hsu, Yau-Heiu ^{[1
]}

Tsai, Hsin-Tzu ^{[1
]}

Liao, Jye-Yann ^{[3
]}

Hu, Chung-Chi ^{[1
]}

机构：

[1] Natl Chung Hsing Univ, Grad Inst Biotechnol, Taichung 402, Taiwan

[2] Acad Sinica, Inst Plant & Microbial Biol, Taipei 115, Taiwan

[3] COA, Agr Res Inst, Plant Pathol Div, Wufeng, Taiwan

来源：

JOURNAL OF VIROLOGICAL METHODS | 2008年 / 147卷 / 02期

关键词：

begomovirus; infectious clone; limited restriction enzyme digestion; rolling circle amplification;

D O I：

10.1016/j.jviromet.2007.10.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (c) 2007 Elsevier B.V. All rights reserved.

引用

页码：355 / 359

页数：5

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