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A Bacillus subtilis spore coat polypeptide gene, cotS
Akiko Abe1, Hidekatsu Koide2, Takeyuki Kohno1
1Faculty of Pharmaceutical Sciences, Setsunan University, 45-1, Nagaotoge-cho, Hirakata, Osaka 573-01, Japan.
Microbiology (Reading, England)
|June 1, 1995
Summary
Researchers identified the Bacillus subtilis cotS gene, crucial for spore coat protein production. This gene, along with orfX, is transcribed during sporulation, suggesting a role in bacterial spore development.
Area of Science:
- Microbiology
- Molecular Biology
- Bacterial Genetics
Background:
- Bacillus subtilis spore coats are complex structures essential for survival.
- Identifying genes involved in spore coat formation is key to understanding bacterial differentiation.
Purpose of the Study:
- To isolate and characterize the gene encoding the Bacillus subtilis spore coat polypeptide Cot40-2.
- To elucidate the genetic organization and transcriptional regulation of the cotS gene.
Main Methods:
- Oligonucleotide-directed gene isolation
- DNA sequencing and ORF analysis
- SDS-PAGE, immunoblotting, and Northern hybridization
- Primer extension analysis for transcriptional start sites
Main Results:
- The Bacillus subtilis cotS gene was isolated and sequenced, encoding a 351-amino acid polypeptide identical to Cot40-2.
- An adjacent open reading frame, orfX, was identified, and both genes are transcribed approximately 5 hours into sporulation.
- Evidence suggests cotS and orfX are part of a multicistronic operon regulated by a sigma K-dependent promoter.
Conclusions:
- The cotS gene product is a major component of the Bacillus subtilis spore coat.
- The cotS and orfX genes are co-transcribed during sporulation, likely under sigma K control.
- Disruption of cotS did not affect growth, sporulation, germination, or chloroform resistance.