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PMID: 8366039 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

New method for gene disruption in Salmonella typhimurium: construction and characterization of an ada-deletion derivative of Salmonella typhimurium TA1535.

Journal of bacteriology ·Vol. 175 ·No. 17 ·1993-09-00 ·Pages 5539-47

Yamada M, Hakura A, Sofuni T, Nohmi T

Abstract

A new method for gene disruption in Salmonella typhimurium was developed. The key steps of this method are to produce restriction fragments with compatible ends, preligate to produce concatemers, and then transform by electrotransformation. We developed and used this method to construct a mutant of S. typhimurium TA1535 in which the resident ada-like (adaST) gene was replaced with a kanamycin resistance gene to produce an adaST-deletion mutant derivative. The S. typhimurium adaST-deletion strain did not exhibit a higher level of mutability upon treatment with N-methyl-N'-nitro-N-nitrosoguanidine than did its wild-type parent strain. However, it did exhibit a higher sensitivity with respect to killing by N-methyl-N'-nitro-N-nitrosoguanidine. The ability of AdaST to function as a transcriptional activator is discussed.

Related Genes
ada
MeSH Terms
Bacterial Proteins/genetics Blotting, Southern Cloning, Molecular DNA Ligases DNA, Bacterial/genetics,metabolism Escherichia coli Proteins Gene Deletion Kanamycin Resistance/genetics Methylation Methyltransferases/genetics Mutagenesis O(6)-Methylguanine-DNA Methyltransferase Restriction Mapping Salmonella typhimurium/genetics Transcription Factors Transformation, Bacterial
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins Transcription Factors Methyltransferases Ada protein, E coli O(6)-Methylguanine-DNA Methyltransferase DNA Ligases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Yamada M
Division of Genetics and Mutagenesis, National Institute of Hygienic Sciences, Tokyo, Japan.
Hakura A
Sofuni T
Nohmi T
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.175.17.5539-5547.1993
Published
1993-09-00
Pages
5539-47
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC206610
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.175.17.5539-5547.1993 PMC Full Text View journal details
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