Sequence and characterization of Bacillus subtilis CheW

D. W. Hanlon, L. M. Marquez-Magana, P. B. Carpenter, M. J. Chamberlin, G. W. Ordal

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)12055-12060
Number of pages6
JournalJournal of Biological Chemistry
Volume267
Issue number17
StatePublished - 1992

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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