| Autor: |
Xu, H. Howard, Trawick, John D., Haselbeck, Robert J., Forsyth, R. Allyn, Yamamoto, Robert T., Archer, Rich, Patterson, Joe, Allen, Molly, Froelich, Jamie M., Taylor, Ian, Nakaji, Danny, Maile, Randy, G. C., Kedar, Pilcher, Marshall, Brown-Driver, Vickie, McCarthy, Melissa, Files, Amy, Robbins, David, King, Paula, Sillaots, Susan, Malone, Cheryl, Zamudio, Carlos S., Roemer, Terry, Wang, Liangsu, Youngman, Philip J., Wall, Daniel |
| Zdroj: |
Antimicrobial Agents and Chemotherapy; September 2010, Vol. 54 Issue: 9 p3659-3670, 12p |
| Abstrakt: |
ABSTRACTThe widespread emergence of antibiotic-resistant bacteria and a lack of new pharmaceutical development have catalyzed a need for new and innovative approaches for antibiotic drug discovery. One bottleneck in antibiotic discovery is the lack of a rapid and comprehensive method to identify compound mode of action (MOA). Since a hallmark of antibiotic action is as an inhibitor of essential cellular targets and processes, we identify a set of 308 essential genes in the clinically important pathogen Staphylococcus aureus. A total of 446 strains differentially expressing these genes were constructed in a comprehensive platform of sensitized and resistant strains. A subset of strains allows either target underexpression or target overexpression by heterologous promoter replacements with a suite of tetracycline-regulatable promoters. A further subset of 236 antisense RNA-expressing clones allows knockdown expression of cognate targets. Knockdown expression confers selective antibiotic hypersensitivity, while target overexpression confers resistance. The antisense strains were configured into a TargetArray in which pools of sensitized strains were challenged in fitness tests. A rapid detection method measures strain responses toward antibiotics. The TargetArray antibiotic fitness test results show mechanistically informative biological fingerprints that allow MOA elucidation. |
| Databáze: |
Supplemental Index |
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