Abstract
A Bacillus subtilis open reading frame (ORF) encoding a predicted polypeptide of 156 amino acids was subcloned and sequenced. The polypeptide was found to be homologous to CheW of Escherichia coli, sharing 28.6% amino acid identity. The ORF was verified by using a bacteriophage T7 expression system in E. coli. The gene was inactivated by insertion of a nonpolor chloramphenicol acetyltransferase cassette in its N-terminal region. In the absence of chemoeffectors, the mutant displayed a smooth swimming bias, with some tumbling. The CheW- mutant was defective on swarm plates but was complemented by a plasmid that expressed wild type CheW. Addition of attractant or repellent to the CheW- mutant resulted in transient smooth swimming or tumbling, respectively. However, capillary assays revealed that chemotaxis was substantially impaired in the mutant strain.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Aspartic Acid / metabolism
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Bacillus subtilis / genetics*
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Bacterial Proteins / genetics*
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Base Sequence
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Blotting, Southern
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Chemotactic Factors / genetics*
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Chloramphenicol O-Acetyltransferase / genetics
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DNA, Bacterial / genetics
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Deoxyglucose / metabolism
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli Proteins*
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Genes, Bacterial
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Genetic Complementation Test
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Molecular Sequence Data
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Open Reading Frames
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Plasmids
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Restriction Mapping
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Sequence Alignment
Substances
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Bacterial Proteins
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CheW protein, E coli
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Chemotactic Factors
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DNA, Bacterial
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Escherichia coli Proteins
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CheW protein, Bacteria
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Aspartic Acid
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Deoxyglucose
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Chloramphenicol O-Acetyltransferase
Associated data
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GENBANK/L01141
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GENBANK/M80251
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GENBANK/M80490
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GENBANK/M80491
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GENBANK/M80492
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GENBANK/S38086
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GENBANK/S96735
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GENBANK/S96741
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GENBANK/S96751
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GENBANK/S96754