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PMID: 2900832 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and expression of a Salmonella enteritidis fimbrin gene in Escherichia coli.

Journal of bacteriology ·Vol. 170 ·No. 9 ·1988-09-00 ·Pages 4216-22

Feutrier J, Kay WW, Trust TJ

Abstract

A gene bank of DNA from a human isolate of Salmonella enteritidis was constructed in the cosmid pHC79 in Escherichia coli HB101. Five clones containing 35- to 45-kilobase inserts of S. enteritidis DNA reacted in colony immunoblot assays with a polyclonal antiserum prepared against purified S. enteritidis fimbriae. Electron microscopy showed that none of the five fimbrin-producing clones produced fimbriae, yet radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis located the 14,400-molecular-weight S. enteritidis in the outer membrane fraction of three of the clones and in the periplasmic fraction of all five clones. By using an oligonucleotide probe homologous to the 5' region of the fimbrin structural gene, the fimbrin gene was located on a 5.3-kilobase HindIII fragment. In vitro transcription-translation analysis verified that this HindIII fragment subcloned into plasmid pTZ18R produced unprocessed S. enteritidis fimbrin of molecular weight 16,400. Dot blot hybridization against a selection of strains of the family Enterobacteriaceae indicated a limited distribution of the S. enteritidis fimbrin gene.

MeSH Terms
Autoradiography Cloning, Molecular Cosmids DNA Restriction Enzymes DNA, Bacterial/genetics Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics Fimbriae, Bacterial/physiology Gene Expression Regulation Genes Genes, Bacterial Humans Immunoassay Membrane Glycoproteins/analysis,genetics Microfilament Proteins Nucleic Acid Hybridization Salmonella enteritidis/analysis,genetics,ultrastructure
Chemicals
DNA, Bacterial Membrane Glycoproteins Microfilament Proteins plastin DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Feutrier J
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Kay W W
Trust T J
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34 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.170.9.4216-4222.1988
Published
1988-09-00
Pages
4216-22
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC211430
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.170.9.4216-4222.1988 PMC Full Text View journal details
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