Mini-Tn7 vectors as genetic tools for gene cloning at a single copy number in an industrially important and phytopathogenic bacteria, Xanthomonas spp. Research letter

Thichakorn Jittawuttipoka, Sarinya Buranajitpakorn, Mayuree Fuangthong, Herbert P. Schweizer, Paiboon Vattanaviboon, Skorn Mongkolsuk

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

Transposon mini-Tn7 vectors insert into the chromosome of several Gram-negative bacteria in a site-specific manner. Here, we showed the application of mini-Tn7 as single copy site-specific integration vector system for Xanthomonas campestris pv. campestris. The transposition of the mini-Tn7 into the bacterial genome was detected at a Tn7 attachment (attTn7) site located downstream of glmS1. Furthermore, using a newly constructed vector pBBR1FLP2 containing the flipase (FLP) recombinase for site-specific excision of the sequence between the FLP recombinase target (FRT) sites, and a sacB counter selection marker, an unmarked mini-Tn7 insertion mutant was created. Mini-Tn7 insertion did not affect bacterial virulence on the tested plant. The mini-Tn7 and FLP-FRT systems also work well in Xanthomonas oryzae pv. oryzae.

Original languageEnglish (US)
Pages (from-to)111-117
Number of pages7
JournalFEMS Microbiology Letters
Volume298
Issue number1
DOIs
StatePublished - Sep 2009
Externally publishedYes

Keywords

  • Mini-Tn7
  • Single copy number gene cloning
  • Unmarked mutation
  • Virulence
  • Xanthomonas

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

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