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Product Name:
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JE2, Transposon Mutant NE1397 (SAUSA300_2166)
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Manufacturer:
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BEI Resources
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Taxonomy:
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Bacteria Classification: Staphylococcaceae, Staphylococcus Species: Staphylococcus aureus Strain: JE2 (also referred to as USA300 JE2) Transposon Mutant: NE1397 Nebraska Transposon Mutant: SAUSA300_2166
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Additional Information:
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In an effort to enhance the research capabilities of the staphylococcal research community, the Center for Staphylococcal Research (CSR) at the University of Nebraska Medical Center has generated the Nebraska Transposon Mutant Library, a collection of sequence-defined transposon (Tn) insertion mutants of S. aureus. This collection contains mutant derivatives of strain USA300 LAC, in which approximately 2,000 non-essential genes have been disrupted by the insertion of the mariner-based transposon bursa aurealis.2 The insertion sites were identified by determining the nucleotide sequences of the junction fragments containing the end of the transposon and the flanking DNA. The gene names and descriptions associated with each of the Tn mutants were obtained from the National Center for Biotechnology Information.
The Nebraska Transposon Mutant Library (NTML) was constructed in the laboratories of Dr. Kenneth Bayles and Dr. Paul Fey at the University of Nebraska Medical Center. Additional information is available at the NTML website.
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Material Provided:
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Each vial contains approximately 0.5 mL of bacterial culture in Tryptic Soy broth containing 5 µg/mL erythromycin supplemented with 10% glycerol.
Note: If homogeneity is required for your intended use, please purify prior to initiating work. It is recommended that the presence and location of the transposon is confirmed. The Center for Staphylococcal Research has designed primers that anneal to the bursa aurealis transposon and are to be used in conjunction with a primer within the gene of interest to generate a PCR product. For transposons in the “plus” orientation, the primer “Upstream” (5’-CTCGATTCTATTAACAAGGG-3’) should be paired with a gene-specific primer. For transposons in the “minus” orientation, the primer “Buster” (5’-GCTTTTTCTAAATGTTTTTTAAGTAAATCAAGTAC-3’) should be paired with a gene-specific primer.
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Packing/Storage:
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NR-47939 was packaged aseptically in cryovials. The product is provided frozen and should be stored at -60°C or colder immediately upon arrival. For long-term storage, the vapor phase of a liquid nitrogen freezer is recommended. Freeze-thaw cycles should be avoided.
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Growth Conditions:
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Media: Tryptic Soy broth containing 5 µg/mL erythromycin or equivalent Tryptic Soy agar containing 5 µg/mL erythromycin or equivalent
Incubation: Temperature: 37°C Atmosphere: Aerobic Propagation: 1. Keep vial frozen until ready for use, then thaw. 2. Transfer the entire thawed aliquot into a single tube of broth. 3. Use several drops of the suspension to inoculate an agar slant and/or plate. 4. Incubate the tube, slant and/or plate at 37°C for 1 day. |
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Disclaimers:
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You are authorized to use this product for research use only. It is not intended for human use. Use of this product is subject to the terms and conditions of the BEI Resources Material Transfer Agreement (MTA). The MTA is available on our Web site at www.beiresources.org. While BEI Resources uses reasonable efforts to include accurate and up-to-date information on this product sheet, neither ATCC® nor the U.S. Government makes any warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. Neither ATCC® nor the U.S. Government warrants that such information has been confirmed to be accurate. This product is sent with the condition that you are responsible for its safe storage, handling, use and disposal. ATCC® and the U.S. Government are not liable for any damages or injuries arising from receipt and/ or use of this product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, the U.S. Government, ATCC®, their suppliers and contributors to BEI Resources are not liable for damages arising from the misidentification or misrepresentation of products. |
References:
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1. Bae, T., et al. “Staphylococcus aureus Virulence Genes Identified by bursa aurealis Mutagenesis and Nematode Killing.” Proc. Natl. Acad. Sci. USA 101 (2004): 12312-12317. PubMed: 15304642.
2. Fey, P. D., et al. “A Genetic Resource for Rapid and Comprehensive Phenotype Screening of Nonessential Staphylococcus aureus Genes.” mBio 4 (2013): e00537-12. PubMed: 23404398.
3. Renna, M. C., et al. “Regulation of the Bacillus subtilis alsS, alsD, and alsR Genes Involved in Post-Exponential-Phase Production of Acetoin.” J. Bacteriol. 175 (1993): 3863-3875. PubMed: 7685336.
4. Yang, S. J., et al. “Characterization of the Staphylococcus aureus CidR Regulon: Elucidation of a Novel Role for Acetoin Metabolism in Cell Death and Lysis.” Mol. Microbiol. 60 (2006): 458-468. PubMed: 16573694.
5. Diep, B. A., et al. “Roles of 34 Virulence Genes in the Evolution of Hospital- and Community-Associated Strains of Methicillin-Resistant Staphylococcus aureus.” J. Infect. Dis. 193 (2006): 1495-1503. PubMed: 16652276.
6. Diekema, D. J., et al. “Continued Emergence of USA300 Methicillin-Resistant Staphylococcus aureus in the United States: Results from a Nationwide Surveillance Study.” Infect. Control Hosp. Epidemiol. 35 (2014): 285-292. PubMed: 24521595.
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Citation:
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Acknowledgment for publications should read "The following reagent was provided by the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA) for distribution through BEI Resources, NIAID, NIH: Staphylococcus aureus subsp. aureus, Strain JE2, Transposon Mutant NE1397 (SAUSA300_2166), NR-47939."
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Biosafety Level:
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2
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories (BMBL). Current Edition. Washington, DC: U.S. Government Printing Office.
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