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Cloning and nucleotide sequence of the type E staphylococcal enterotoxin gene
J L Couch1, M T Soltis, M J Betley
1Department of Bacteriology, University of Wisconsin-Madison 53706.
Journal of Bacteriology
|July 1, 1988
Summary
Researchers cloned the staphylococcal enterotoxin E (entE) gene into E. coli, determining its sequence and protein structure. This study reveals entE
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Staphylococcal enterotoxins are potent exotoxins produced by Staphylococcus aureus.
- Understanding the genetic basis of enterotoxin production is crucial for food safety and public health.
Purpose of the Study:
- To clone and characterize the staphylococcal enterotoxin type E (entE) gene.
- To determine the nucleotide and amino acid sequences of staphylococcal enterotoxin E.
- To compare the entE gene and its product with other staphylococcal enterotoxins.
Main Methods:
- Gene cloning using plasmid vector pBR322.
- Bacterial transformation of Escherichia coli.
- DNA sequencing and N-terminal amino acid sequencing.
- Sequence homology analysis.
Main Results:
- The staphylococcal enterotoxin type E (entE) gene was successfully cloned and expressed in E. coli.
- The complete nucleotide sequence of entE (771 base pairs) and the N-terminal amino acid sequence of mature staphylococcal enterotoxin E were determined.
- The entE gene encodes a 29,358 Da precursor protein, processed to a 26,425 Da mature extracellular form.
- Staphylococcal enterotoxins E and A show significant sequence homology (84% nucleotide, 83% amino acid).
Conclusions:
- The entE gene sequence and protein structure have been elucidated.
- Staphylococcal enterotoxin E is closely related to staphylococcal enterotoxin A at both the nucleotide and amino acid levels.
- This genetic information contributes to understanding the diversity and evolution of staphylococcal enterotoxins.