In vivo excision, cloning, and broad-host-range transfer of large bacterial DNA segments using VEX-Capture.

James W. Wilson, Cheryl Nickerson

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The performance of many bacterial genetic experiments would benefit from a convenient method to clone large sets of genes (20-100+ kb) and transfer these genes to a wide range of other bacterial recipients. The VEX-Capture technique allows such large genomic segments to be cloned in vivo onto a broad-host-range IncP plasmid that is able to self-transfer to a wide variety of Gram-negative bacteria. The advantages of VEX-Capture are its efficiency, specificity, and use of common molecular biological techniques that do not require non-standard equipment and are easily applicable to many types of bacterial species. Here, we describe the VEX-Capture experimental protocol using Salmonella typhimurium as the source of the target DNA segment.

Original languageEnglish (US)
Pages (from-to)105-118
Number of pages14
JournalMethods in molecular biology (Clifton, N.J.)
Volume394
DOIs
StatePublished - 2007

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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