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J Microbiol Methods. 2009 Dec;79(3):314-20. doi: 10.1016/j.mimet.2009.09.025. Epub 2009 Oct 13.

Changes of physiological and biochemical properties of Salmonella enterica serovar Typhimurium by deletion of cpxR and lon genes using allelic exchange method.

Author information

1
Division of Biological Sciences, Pusan National University, Busan 609-735, South Korea.

Abstract

To construct a novel Salmonella attenuated live vaccine, the cpxR and lon genes were deleted from a wild-type Salmonella enterica serovar Typhimurium (S. Typhimurium) using allelic exchange method, resulting in S. Typhimurium CK31 (DeltacpxR), CK38 (Deltalon), and CK111 (DeltacpxR/lon). These mutated strains were grown normally, as was the wild-type strain. The biochemical properties of the mutants remained highly similar to those of the wild-type. In comparison with the wild-type, 1.5 to 3.3-fold increases of fimbrial products such as Agf, Fim, and Pef fimbria in the mutants CK31, CK38, and CK111 were observed by using a transmission electron microscope and dot blotting. Furthermore, CK38 and CK111 morphologically appeared elongated in shape and produced 2.0- and 3.2-fold increases, respectively, of capsular polysaccharide, which is a major antigenic component. Approximately 10(4)-fold attenuation assessed by analysis of LD(50) of BALB/c mouse was observed by deleting the lon/cpxR (CK111) genes. This result indicated that deletion of lon and cpxR genes induced significant attenuation.

PMID:
19833153
DOI:
10.1016/j.mimet.2009.09.025
[Indexed for MEDLINE]

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