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FEMS Microbiol Lett. 2005 Apr 15;245(2):315-9.

Efficient gene inactivation in Bacillus anthracis.

Author information

1
Center for Pharmaceutical Biotechnology, University of Illinois, M/C 870, 900 S. Ashland Ave., Chicago, IL 60607, USA. shatalin@uic.edu

Abstract

A procedure for high-efficiency gene inactivation in Bacillus anthracis has been developed. It is based on a highly temperature-sensitive plasmid vector carrying kanamycin resistance cassette surrounded by DNA fragments flanking the desired insertion site. The approach was tested by constructing glutamate racemase E1 (racE1), glutamate racemase E2 (racE2) and comEC knock-out mutants of B. anthracis strain DeltaANR. Allelic replacements were observed at high frequencies, ranging from approximately 0.5% for racE2 up to 50% for racE1 and comEC. The system can be used for genetic validation of potential drug targets.

PMID:
15837388
DOI:
10.1016/j.femsle.2005.03.029
[Indexed for MEDLINE]
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