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J Microbiol Methods. 2010 Sep;82(3):296-300. doi: 10.1016/j.mimet.2010.07.002. Epub 2010 Jul 16.

Mini-Tn7 vectors as genetic tools for single copy gene cloning in Acinetobacter baumannii.

Author information

1
Faculty of Health Sciences, University of Ontario Institute of Technology, Oshawa, Ontario, Canada. Ayush.Kumar@uoit.ca

Abstract

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-Tn7-based single copy insertion system in A. baumannii. Mini-Tn7 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-Tn7 in A. baumannii and demonstrated application of this useful cloning tool by inserting the gfp gene into the chromosome. Our work shows that mini-Tn7 elements are useful tools for genetic studies in this important pathogen.

PMID:
20638421
DOI:
10.1016/j.mimet.2010.07.002
[Indexed for MEDLINE]

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