TY - JOUR
T1 - Mini-Tn7 vectors as genetic tools for single copy gene cloning in Acinetobacter baumannii
AU - Kumar, Ayush
AU - Dalton, Courtney
AU - Cortez-Cordova, Jenny
AU - Schweizer, Herbert P.
N1 - Funding Information:
This work was supported by grants from the University of Ontario Institute of Technology and the Canada Foundation for Innovation — Leaders Opportunities Fund to AK and those from a Colorado State University Infectious Diseases Supercluster award to HPS. CD is a recipient of UOIT-STAR award. Authors acknowledge the help of Alan Schenkel, Department of Microbiology, Immunology, and Pathology, Colorado State University with fluorescent microscopy.
PY - 2010/9
Y1 - 2010/9
N2 - Acinetobacter baumannii is an emerging pathogen that causes serious infections with high mortality rates in immunocompromised individuals. Genetic manipulations in this medically-relevant pathogen are limited by the paucity of molecular tools. In this study, we show the application of the mini-Tn. 7-based single copy insertion system in A. baumannii. Mini-Tn. 7 elements are known to integrate at a naturally evolved, therefore presumably neutral location (intergenic region) downstream of the glmS gene (glucosamine-fructose-6-phosphate aminotransferase) in Gram-negative bacteria. We identified the site of insertion of mini-Tn. 7 in A. baumannii and demonstrated application of this useful cloning tool by inserting the gfp gene into the chromosome. Our work shows that mini-Tn 7 elements are useful tools for genetic studies in this important pathogen.
AB - Acinetobacter baumannii is an emerging pathogen that causes serious infections with high mortality rates in immunocompromised individuals. Genetic manipulations in this medically-relevant pathogen are limited by the paucity of molecular tools. In this study, we show the application of the mini-Tn. 7-based single copy insertion system in A. baumannii. Mini-Tn. 7 elements are known to integrate at a naturally evolved, therefore presumably neutral location (intergenic region) downstream of the glmS gene (glucosamine-fructose-6-phosphate aminotransferase) in Gram-negative bacteria. We identified the site of insertion of mini-Tn. 7 in A. baumannii and demonstrated application of this useful cloning tool by inserting the gfp gene into the chromosome. Our work shows that mini-Tn 7 elements are useful tools for genetic studies in this important pathogen.
KW - A. baumannii
KW - Mini-Tn7
KW - Single-copy gene cloning
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U2 - 10.1016/j.mimet.2010.07.002
DO - 10.1016/j.mimet.2010.07.002
M3 - Article
C2 - 20638421
AN - SCOPUS:77955660026
SN - 0167-7012
VL - 82
SP - 296
EP - 300
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
IS - 3
ER -