Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression

Robert A. Edwards; Linda H. Keller; Dieter M. Schifferli

doi:10.1016/S0378-1119(97)00619-7

Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression

Robert A. Edwards, Linda H. Keller, Dieter M. Schifferli

Research output: Contribution to journal › Article › peer-review

480 Citations (Scopus)

Abstract

A series of vectors has been developed to provide improved positive and negative selection for allelic exchange. Based on homologous regions of DNA ranging in size from less than 200 bp to over 1 kb, we have successfully used these new plasmids to introduce or remove markers in chromosomal or plasmid DNA. Wild type fimbria genes were replaced both in Salmonella enteritidis (sefA, agfA and fimC) and Escherichia coli (fasA and fasH). Regulation of 987P fimbriation could be identified after replacement of fasA and fasH with allelic reporter fusions. The expression of fasA but not fasH is dependent upon the osmolarity of the growth medium in an HNS-dependent manner, but unlike some other fimbrial systems expression is not dependent on the exogenous iron concentration.

Original language	English
Pages (from-to)	149-157
Number of pages	9
Journal	Gene
Volume	207
Issue number	2
DOIs	https://doi.org/10.1016/S0378-1119(97)00619-7
Publication status	Published - 30 Jan 1998
Externally published	Yes

Keywords

987P
Fimbriae
hns
Osmolarity
Pili
sacB

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abstract = "A series of vectors has been developed to provide improved positive and negative selection for allelic exchange. Based on homologous regions of DNA ranging in size from less than 200 bp to over 1 kb, we have successfully used these new plasmids to introduce or remove markers in chromosomal or plasmid DNA. Wild type fimbria genes were replaced both in Salmonella enteritidis (sefA, agfA and fimC) and Escherichia coli (fasA and fasH). Regulation of 987P fimbriation could be identified after replacement of fasA and fasH with allelic reporter fusions. The expression of fasA but not fasH is dependent upon the osmolarity of the growth medium in an HNS-dependent manner, but unlike some other fimbrial systems expression is not dependent on the exogenous iron concentration.",

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AU - Edwards, Robert A.

AU - Keller, Linda H.

AU - Schifferli, Dieter M.

PY - 1998/1/30

Y1 - 1998/1/30

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KW - 987P

KW - Fimbriae

KW - hns

KW - Osmolarity

KW - Pili

KW - sacB

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Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression

Abstract

Keywords

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