New Vector Tools with a Hygromycin Resistance Marker for Use with Opportunistic Pathogens
Eric J. Kalivoda,Eric J. Kalivoda,Joseph Horzempa,Nicholas A. Stella,Aroba Sadaf,Regis P. Kowalski,Gerard J. Nau,Robert M. Q. Shanks +7 more
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TLDR
A diverse set of vectors are modified to include a broadly expressed hygromycin resistance (HmR) marker and each vector has an origin of transfer for conjugation, and is able to replicate in Saccharomyces cerevisiae to take advantage of the powerful yeast recombineering system.Abstract:
The ability of many bacterial strains to tolerate antibiotics can limit the number of molecular tools available for research of these organisms. To help address this problem, we have modified a diverse set of vectors to include a broadly expressed hygromycin resistance (HmR) marker. Hygromycin B is an aminoglycoside antibiotic not used to treat infections in humans and has antimicrobial activity against a wide range of microorganisms. Vectors with four replication origins are represented, with potential applications including general cloning, allelic replacement, and transcriptional analysis. We show that vectors with the broad host range pBBR1-replicon conferred HmR to Achromobacter xylosoxidans, Acinetobacter baumannii, Pseudomonas aeruginosa, and Serratia marcescens, and a pC194-based vector was able to confer HmR to Francisella tularensis. We also used a subset of these plasmids to manipulate the genome of S. marcescens. Each vector has an origin of transfer for conjugation, and is also able to replicate in Saccharomyces cerevisiae to take advantage of the powerful yeast recombineering system.read more
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AB5075, a Highly Virulent Isolate of Acinetobacter baumannii, as a Model Strain for the Evaluation of Pathogenesis and Antimicrobial Treatments
Anna C. Jacobs,Mitchell G. Thompson,Chad C. Black,Jennifer L. Kessler,Lily P. Clark,Christin N. McQueary,Hanan Gancz,Brendan W. Corey,Jay K. Moon,Yuanzheng Si,Matthew T. Owen,Justin D. Hallock,Yoon I. Kwak,Amy Summers,Charles Z. Li,David A. Rasko,William F. Penwell,Cary L. Honnold,Matthew C. Wise,Paige E. Waterman,Emil Lesho,Rena L. Stewart,Luis A. Actis,Thomas J. Palys,David Craft,Daniel V. Zurawski +25 more
TL;DR: It is proposed that AB5075 can serve as a model strain for A. baumannii pathogenesis due to its relatively recent isolation, multidrug resistance, reproducible virulence in animal models, and genetic tractability.
Journal ArticleDOI
Antibiotic failure mediated by a resistant subpopulation in Enterobacter cloacae
Victor I. Band,Emily K. Crispell,Brooke A. Napier,Carmen M. Herrera,Greg K. Tharp,Kranthi Vavikolanu,Jan Pohl,Timothy D. Read,Steven E. Bosinger,Steven E. Bosinger,M. Stephen Trent,Eileen M. Burd,David S. Weiss +12 more
TL;DR: The ability of low-frequency bacterial subpopulations to contribute to clinically relevant antibiotic resistance is demonstrated, elucidating an enigmatic cause of antibiotic treatment failure and highlighting the critical need for more sensitive diagnostics.
Journal ArticleDOI
Contribution of β-Lactamases and Porin Proteins OmpK35 and OmpK36 to Carbapenem Resistance in Clinical Isolates of KPC-2-Producing Klebsiella pneumoniae
Ying Zhang,Xiaofei Jiang,Xiaofei Jiang,Yan-yan Wang,Gang Li,Yueru Tian,Hong Liu,Fuqi Ai,Yiming Ma,Bei Wang,Feiyi Ruan,Kumar Rajakumar,Kumar Rajakumar +12 more
TL;DR: Fifty-seven carbapenem-resistant Klebsiella pneumoniae isolates belonging to ST11, ST423, and two other sequence types were studied and blaKPC-2, blaTEM-1, andBlaCTX-M-14 demonstrated varied porin expression.
Journal ArticleDOI
IS5 element integration, a novel mechanism for rapid in vivo emergence of tigecycline nonsusceptibility in Klebsiella pneumoniae.
Lindsey Nielsen,Erik Snesrud,Fatma Onmus-Leone,Yoon I. Kwak,Ricardo Aviles,Eric Steele,Deena Sutter,Paige E. Waterman,Emil Lesho +8 more
TL;DR: This is the first report to propose a function for kpgABC and identify an insertion element whose presence correlated with the in vivo development of tigecycline nonsusceptibility in K. pneumoniae.
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Identification of SlpB, a Cytotoxic Protease from Serratia marcescens
TL;DR: Genetic data introduce SlpB as a new cytotoxic protease from S. marcescens as well as indicating that the type I secretion system gene, lipD, is required forSlpB secretion.
References
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Journal Article
A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
Journal ArticleDOI
STUDIES ON LYSOGENESIS I.: The Mode of Phage Liberation by Lysogenic Escherichia coli
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