A Tn7-based broad-range bacterial cloning and expression system

Kyoung Hee Choi, Jared B. Gaynor, Kimberly G. White, Carolina Lopez, Catharine M. Bosio, Rox Ann R. Karkhoff-Schweizer, Herbert P. Schweizer

Research output: Contribution to journalArticlepeer-review

502 Scopus citations

Abstract

For many bacteria, cloning and expression systems are either scarce or nonexistent. We constructed several mini-Tn7 vectors and evaluated their potential as broad-range cloning and expression systems. In bacteria with a single chromosome, including Pseudomonas aeruginosa, Pseudomonas putida and Yersinia pestis, and in the presence of a helper plasmid encoding the site-specific transposition pathway, site- and orientation-specific Tn7 insertions occurred at a single attTn7 site downstream of the glmS gene. Burkholderia thailandensis contains two chromosomes, each containing a glmS gene and an attTn7 site. The Tn7 system allows engineering of diverse genetic traits into bacteria, as demonstrated by complementing a biofilm-growth defect of P. aeruginosa, establishing expression systems in P. aeruginosa and P. putida, and 'GFP-tagging' Y. pestis. This system will thus have widespread biomedical and environmental applications, especially in environments where plasmids and antibiotic selection are not feasible, namely in plant and animal models or biofilms.

Original languageEnglish (US)
Pages (from-to)443-448
Number of pages6
JournalNature Methods
Volume2
Issue number6
DOIs
StatePublished - Jun 2005
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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