Genome engineering in Bacillus anthracis using Cre recombinase.

ABSTRACT: Genome engineering is a powerful method for the study of bacterial virulence. With the availability of the complete genomic sequence of Bacillus anthracis, it is now possible to inactivate or delete selected genes of interest. However, many current methods for disrupting or deleting more than one gene require use of multiple antibiotic resistance determinants. In this report we used an approach that temporarily inserts an antibiotic resistance marker into a selected region of the genome and subsequently removes it, leaving the target region (a single gene or a larger genomic segment) permanently mutated. For this purpose, a spectinomycin resistance cassette flanked by bacteriophage P1 loxP sites oriented as direct repeats was inserted within a selected gene. After identification of strains

SUBMITTER: Pomerantsev AP 

PROVIDER: S-EPMC1346652 | biostudies-literature | 2006 Jan

REPOSITORIES: biostudies-literature