Identification of Essential Genes in Staphylococcus aureus by Construction and Screening of Conditional Mutant Library

2008 ◽  
pp. 297-305 ◽  
Author(s):  
Dezhong Yin ◽  
Yinduo Ji
Keyword(s):  
Staphylococcus Aureus ◽  
Essential Genes ◽  
Mutant Library ◽  
Conditional Mutant

scholarly journals Shuttle Vectors Derived from pN315 for Study of Essential Genes in Staphylococcus aureus

2010 ◽  
Vol 33 (2) ◽  
pp. 198-203 ◽  
Author(s):  
Miki Matsuo ◽  
Kenji Kurokawa ◽  
Bok-Luel Lee ◽  
Kazuhisa Sekimizu
Keyword(s):  
Staphylococcus Aureus ◽  
Essential Genes ◽  
Shuttle Vectors

scholarly journals B urkholderia cenocepacia conditional growth mutant library created by random promoter replacement of essential genes

2013 ◽  
Vol 2 (2) ◽  
pp. 243-258 ◽  
Author(s):  
Ruhi A. M. Bloodworth ◽  
April S. Gislason ◽  
Silvia T. Cardona
Keyword(s):  
Essential Genes ◽  
Mutant Library ◽  
Promoter Replacement

scholarly journals A genome-wide strategy for the identification of essential genes in Staphylococcus aureus

2002 ◽  
Vol 43 (6) ◽  
pp. 1387-1400 ◽  
Author(s):  
R. Allyn Forsyth ◽  
Robert J. Haselbeck ◽  
Kari L. Ohlsen ◽  
Robert T. Yamamoto ◽  
Howard Xu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Essential Genes ◽  
Genome Wide ◽  
A Genome

Construction of a Sequence-Defined Transposon Mutant Library in Staphylococcus aureus

2019 ◽  
pp. 29-37 ◽  
Author(s):  
Jennifer L. Endres ◽  
Vijaya Kumar Yajjala ◽  
Paul D. Fey ◽  
Kenneth W. Bayles
Keyword(s):  
Staphylococcus Aureus ◽  
Mutant Library

Generation of a Transposon Mutant Library in Staphylococcus aureus and Staphylococcus epidermidis Using bursa aurealis

2014 ◽  
pp. 103-110 ◽  
Author(s):  
Vijaya Kumar Yajjala ◽  
Todd J. Widhelm ◽  
Jennifer L. Endres ◽  
Paul D. Fey ◽  
Kenneth W. Bayles
Keyword(s):  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Mutant Library

scholarly journals Genes Contributing to Staphylococcus aureus Fitness in Abscess- and Infection-Related Ecologies

mBio ◽  
2014 ◽  
Vol 5 (5) ◽  
Author(s):  
Michael D. Valentino ◽  
Lucy Foulston ◽  
Ama Sadaka ◽  
Veronica N. Kos ◽  
Regis A. Villet ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
New Drugs ◽  
Insertion Mutant ◽  
Mutant Library ◽  
Entire Genome ◽  
Antibiotic Resistant ◽  
Content Type ◽  
Transposon Insertion ◽  
Genes Encoding ◽  
Novel Targets

ABSTRACTStaphylococcus aureusis a leading cause of both community- and hospital-acquired infections that are increasingly antibiotic resistant. The emergence ofS. aureusresistance to even last-line antibiotics heightens the need for the development of new drugs with novel targets. We generated a highly saturated transposon insertion mutant library in the genome ofS. aureusand used Tn-seq analysis to probe the entire genome, with unprecedented resolution and sensitivity, for genes of importance in infection. We further identified genes contributing to fitness in various infected compartments (blood and ocular fluids) and compared them to genes required for growth in rich medium. This resulted in the identification of 426 genes that were important forS. aureusfitness during growth in infection models, including 71 genes that could be considered essential for survival specifically during infection. These findings highlight novel as well as previously known genes encoding virulence traits and metabolic pathways important forS. aureusproliferation at sites of infection, which may represent new therapeutic targets.IMPORTANCEStaphylococcus aureuscontinues to be a leading cause of antibiotic-resistant community and nosocomial infection. With the bacterium’s acquisition of resistance to methicillin and, more recently, vancomycin, the need for the development of new drugs with novel targets is urgent. Applying a highly saturated Tn-seq mutant library to analyze fitness and growth requirements in a murine abscess and in various infection-relevant fluids, we identifiedS. aureustraits that enable it to survive and proliferate during infection. This identifies potential new targeting opportunities for the development of novel therapeutics.

scholarly journals Defined Mutant Library Sequencing (DML-Seq) for Identification of Conditional Essential Genes

BIO-PROTOCOL ◽  
2021 ◽  
Vol 11 (5) ◽  
Author(s):  
Shuai Shao ◽  
Lifan Wei ◽  
Feng Xia ◽  
Yuanxing Zhang ◽  
Qiyao Wang
Keyword(s):  
Essential Genes ◽  
Mutant Library

scholarly journals Essential genes of the macrophage response to Staphylococcus aureus exposure

2018 ◽  
Vol 23 (1) ◽  
Author(s):  
Aixia Sun ◽  
Hongwei Zhang ◽  
Feng Pang ◽  
Guifen Niu ◽  
Jianzhong Chen ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Essential Genes ◽  
Macrophage Response

scholarly journals Identification of a Novel LysR-Type Transcriptional Regulator in Staphylococcus aureus That Is Crucial for Secondary Tissue Colonization during Metastatic Bloodstream Infection

mBio ◽  
2020 ◽  
Vol 11 (4) ◽  
Author(s):  
Michaela Groma ◽  
Sarah A. Horst ◽  
Sudip Das ◽  
Bruno Huettel ◽  
Maximilian Klepsch ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bloodstream Infection ◽  
Murine Model ◽  
Bacterial Colonization ◽  
Critical Role ◽  
Transcriptional Regulator ◽  
Metabolic Adaptation ◽  
Mutant Library ◽  
Copper Transporter ◽  
Content Type

ABSTRACT Staphylococcus aureus is a common cause of bacteremia that can lead to severe complications once the bacteria exit the bloodstream and establish infection in secondary organs. Despite its clinical relevance, little is known about the bacterial factors facilitating the development of these metastatic infections. Here, we used an S. aureus transposon mutant library coupled to transposon insertion sequencing (Tn-Seq) to identify genes that are critical for efficient bacterial colonization of secondary organs in a murine model of metastatic bloodstream infection. Our transposon screen identified a LysR-type transcriptional regulator (LTTR), which was required for efficient colonization of secondary organs such as the kidneys in infected mice. The critical role of LTTR in secondary organ colonization was confirmed using an isogenic mutant deficient in the expression of LTTR. To identify the set of genes controlled by LTTR, we used an S. aureus strain carrying the LTTR gene in an inducible expression plasmid. Gene expression analysis upon induction of LTTR showed increased transcription of genes involved in branched-chain amino acid biosynthesis, a methionine sulfoxide reductase, and a copper transporter as well as decreased transcription of genes coding for urease and components of pyrimidine nucleotides. Furthermore, we show that transcription of LTTR is repressed by glucose, is induced under microaerobic conditions, and required trace amounts of copper ions. Our data thus pinpoints LTTR as an important element that enables a rapid adaptation of S. aureus to the changing host microenvironment. IMPORTANCE Staphylococcus aureus is an important pathogen that can disseminate via the bloodstream and establish metastatic infections in distant organs. To achieve a better understanding of the bacterial factors facilitating the development of these metastatic infections, we used in this study a Staphylococcus aureus transposon mutant library in a murine model of intravenous infection, where bacteria first colonize the liver as the primary infection site and subsequently progress to secondary sites such as the kidney and bones. We identified a novel LysR-type transcriptional regulator (LTTR), which was specifically required by S. aureus for efficient colonization of secondary organs. We also determined the transcriptional activation as well as the regulon of LTTR, which suggests that this regulator is involved in the metabolic adaptation of S. aureus to the host microenvironment found in secondary infection sites.

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