TY - JOUR
T1 - Sequence and characterization of Bacillus subtilis CheW
AU - Hanlon, David W.
AU - Marquez-Magańa, Leticia M.
AU - Carpenter, Phillip B.
AU - Chamberlin, Michael J.
AU - Ordal, George W.
N1 - Copyright:
Copyright 2004 Elsevier B.V., All rights reserved.
PY - 1992/6/15
Y1 - 1992/6/15
N2 - 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 substan-tially impaired in the mutant strain.
AB - 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 substan-tially impaired in the mutant strain.
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M3 - Article
C2 - 1601874
AN - SCOPUS:0026684614
SN - 0021-9258
VL - 267
SP - 12055
EP - 12060
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 17
ER -