TY - JOUR
T1 - A Tn7-based broad-range bacterial cloning and expression system
AU - Choi, Kyoung Hee
AU - Gaynor, Jared B.
AU - White, Kimberly G.
AU - Lopez, Carolina
AU - Bosio, Catharine M.
AU - Karkhoff-Schweizer, Rox Ann R.
AU - Schweizer, Herbert P.
N1 - Funding Information:
H.P.S. was supported by National Institutes of Health grant AI058141. We thank K. Giesler and K. Quinn for their contributions to this project, and R. Gillis (University of Rochester) for assisting with biofilm experiments.
PY - 2005/6
Y1 - 2005/6
N2 - 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.
AB - 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.
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U2 - 10.1038/nmeth765
DO - 10.1038/nmeth765
M3 - Article
C2 - 15908923
AN - SCOPUS:21444457864
SN - 1548-7091
VL - 2
SP - 443
EP - 448
JO - Nature Methods
JF - Nature Methods
IS - 6
ER -