Cloning of Circular DNAs from Microorganisms Using a Novel Plasmid Capture System
Plasmid capture system (PCS) facilitates cloning and manipulation of circular double-stranded DNA. We recently developed an improved PCS (PCS-LZ) to clone relatively large DNA molecules of 30-150 kb. The PCS-LZ donor consists of a mini-F replicon and a kanamycin resistance marker between Tn7 left an...
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Published in | Molecular biotechnology Vol. 44; no. 2; pp. 120 - 126 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
New York
New York : Humana Press Inc
01.02.2010
Humana Press Inc Springer Springer Nature B.V |
Subjects | |
Online Access | Get full text |
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Summary: | Plasmid capture system (PCS) facilitates cloning and manipulation of circular double-stranded DNA. We recently developed an improved PCS (PCS-LZ) to clone relatively large DNA molecules of 30-150 kb. The PCS-LZ donor consists of a mini-F replicon and a kanamycin resistance marker between Tn7 left and Tn7 right ends. Both the replicon and marker gene of the PCS-LZ donor are transferred into target plasmid DNAs by in vitro transposition, followed by replication in E. coli. Colonies are tested for lacZ expression by blue/white screening. Circular DNAs were obtained from plasmids of Bacillus thuringiensis, genome segments of Cotesia glomerata bracovirus and polymorphic genomes of Autographa californica nucleopolyhedrovirus. PCS-LZ is a powerful tool for use in genomic analysis and mutagenesis in microorganisms including invertebrate pathogens. |
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Bibliography: | http://dx.doi.org/10.1007/s12033-009-9215-3 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1073-6085 1559-0305 |
DOI: | 10.1007/s12033-009-9215-3 |