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

Ayush Kumar; Courtney Dalton; Jenny Cortez-Cordova; Herbert P. Schweizer

doi:10.1016/j.mimet.2010.07.002

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

Ayush Kumar
, Courtney Dalton
, Jenny Cortez-Cordova
, Herbert P. Schweizer

Research output: Contribution to journal › Article › peer-review

55 Scopus citations

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-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.

Original language	English (US)
Pages (from-to)	296-300
Number of pages	5
Journal	Journal of Microbiological Methods
Volume	82
Issue number	3
DOIs	https://doi.org/10.1016/j.mimet.2010.07.002
State	Published - Sep 2010
Externally published	Yes

Keywords

A. baumannii
Mini-Tn7
Single-copy gene cloning

ASJC Scopus subject areas

Microbiology
Molecular Biology
Microbiology (medical)

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10.1016/j.mimet.2010.07.002

Cite this

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title = "Mini-Tn7 vectors as genetic tools for single copy gene cloning in Acinetobacter baumannii",

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-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.",

keywords = "A. baumannii, Mini-Tn7, Single-copy gene cloning",

author = "Ayush Kumar and Courtney Dalton and Jenny Cortez-Cordova and Schweizer, \{Herbert P.\}",

note = "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.",

year = "2010",

month = sep,

doi = "10.1016/j.mimet.2010.07.002",

language = "English (US)",

volume = "82",

pages = "296--300",

journal = "Journal of Microbiological Methods",

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AU - Kumar, Ayush

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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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DO - 10.1016/j.mimet.2010.07.002

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AN - SCOPUS:77955660026

SN - 0167-7012

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SP - 296

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JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

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ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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