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

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 r...

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Published in	Journal of virological methods Vol. 147; no. 2; pp. 355 - 359
Main Authors	Wu, Chia-Ying, Lai, Yi-Chin, Lin, Na-Sheng, Hsu, Yau-Heiu, Tsai, Hsin-Tzu, Liao, Jye-Yann, Hu, Chung-Chi
Format	Journal Article
Language	English
Published	London Elsevier B.V 01.02.2008 Amsterdam Elsevier New York, NY
Subjects	Ageratum yellow vein virus Begomovirus Begomovirus - genetics Biological and medical sciences Cloning, Molecular - methods DNA Restriction Enzymes - metabolism Fundamental and applied biological sciences. Psychology Genome, Viral Infectious clone Limited restriction enzyme digestion Microbiology Plant Diseases - virology Rolling circle amplification Squash leaf curl virus Techniques used in virology Tomato leaf curl virus Virology Infectious clone Limited restriction enzyme digestion Rolling circle amplification Begomovirus Plant pathogen Microbiology Enzyme Method Virology Virus Amplification Infection Geminiviridae
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Abstract	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.
AbstractList	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.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. 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.
Author	Lai, Yi-Chin Tsai, Hsin-Tzu Hsu, Yau-Heiu Hu, Chung-Chi Wu, Chia-Ying Liao, Jye-Yann Lin, Na-Sheng
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Keywords	Infectious clone Limited restriction enzyme digestion Rolling circle amplification Begomovirus Plant pathogen Microbiology Enzyme Method Virology Virus Amplification Infection Geminiviridae
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Snippet	Most infectious clones of geminiviruses consist of (partial) tandem repeats of viral genomes in the vectors, which usually involve tedious, multi-step...
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SubjectTerms	Ageratum yellow vein virus Begomovirus Begomovirus - genetics Biological and medical sciences Cloning, Molecular - methods DNA Restriction Enzymes - metabolism Fundamental and applied biological sciences. Psychology Genome, Viral Infectious clone Limited restriction enzyme digestion Microbiology Plant Diseases - virology Rolling circle amplification Squash leaf curl virus Techniques used in virology Tomato leaf curl virus Virology
Title	A simplified method of constructing infectious clones of begomovirus employing limited restriction enzyme digestion of products of rolling circle amplification
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