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PMID: 3898073 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Molecular cloning of staphylococcal enterotoxin B gene in Escherichia coli and Staphylococcus aureus.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 17 ·1985-09-00 ·Pages 5850-4

Ranelli DM, Jones CL, Johns MB, Mussey GJ, Khan SA

Abstract

We have cloned the Staphylococcus aureus entB gene in Escherichia coli, using pBR322 as the vector plasmid; however, no detectable staphylococcal enterotoxin B (SEB) was produced by the E. coli clones. When the entB gene was placed downstream from the strong lambda phage promoter, PR, SEB was synthesized at readily detectable levels in E. coli. Interestingly, mature SEB was almost exclusively present in the cytoplasmic fraction. The SEB precursor was found associated with the cell membrane. The entB gene was introduced back into S. aureus, and the clones were shown to produce SEB. The entB gene has been located to a 2.1-kilobase-pair region. Maxam-Gilbert sequencing of a part of the entB gene yielded a DNA sequence that corresponds to the known amino acid sequence of SEB. Southern hybridization experiments showed that the entB gene was present on identical restriction fragments in the chromosomes of SEB-producer strains. The entB gene is absent from SEB-nonproducer strains.

MeSH Terms
Chromosomes, Bacterial/ultrastructure Cloning, Molecular DNA, Bacterial/genetics Enterotoxins/genetics Escherichia coli/genetics Gene Expression Regulation Genes, Bacterial Plasmids Staphylococcus aureus/genetics
Chemicals
DNA, Bacterial Enterotoxins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ranelli D M
Jones C L
Johns M B
Mussey G J
Khan S A
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43 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.82.17.5850
Published
1985-09-00
Pages
5850-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC390651
Subset
IM
Grants
NIAID NIH HHS · AI-19783 · United States
Databases
GENBANK
M11118
External Links
PubMed source DOI: 10.1073/pnas.82.17.5850 PMC Full Text View journal details
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