Bacillus subtilis mutant deficient in the major autolytic amidase and glucosaminidase is impaired in motility.
The purified autolytic endo-beta-N-acetylglucosaminidase of Bacillus subtilis AC327 was cleaved with cyanogen bromide, and the N-terminal amino acid sequence of one of the peptide fragments was determined. Then, a DNA fragment containing a part of the glucosaminidase gene was cloned into Escherichia coli JM109 using synthetic oligonucleotides as probes whose sequences had been deduced from the N-terminal amino acid sequence. Zymographic analysis showed that the resultant glucosaminidase-deficient strain lacked a 35-kDa lytic band in addition to a 90-kDa lytic one corresponding to the glucosaminidase. A double mutant strain deficient in the major two autolysins (amidase and glucosaminidase) exhibited greatly impaired motility on a swarm plate whereas the single mutant strains were motile.
Rashid
M H
MH
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan.
Kuroda
A
A
Sekiguchi
J
J
D000118:Acetylglucosaminidase / Q000235:genetics*; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002465:Cell Movement / Q000502:physiology; D004269:DNA, Bacterial / Q000235:genetics; D006596:Hexosaminidases / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes / Q000235:genetics
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D006596:Hexosaminidases; D000118:Acetylglucosaminidase