Molecular cloning of a gene affecting the autolysin level and flagellation in Bacillus subtilis

J Gen Microbiol. 1988 Jun;134(6):1611-21. doi: 10.1099/00221287-134-6-1611.

Abstract

A 2.8 kb PstI fragment of Bacillus subtilis 168W DNA has been cloned into Escherichia coli HB101 and B. subtilis AG5 using pAC3 as a shuttle plasmid. The new plasmid (pBRG1), of 10.2 kb, complemented flaD mutations which show reduced production of autolysin(s), filamentation and non-motility (deficiency of flagella). Deletion experiments showed that the suppressive gene is located between the HindIII and XbaI sites (1.0 kb apart) in pBRG1. The integration of a plasmid having chloramphenicol resistance closely linked to the flaD gene into the B. subtilis AC703 chromosome and its genetic analysis indicated that the cloned fragment contained the flaD gene itself. A high-copy-number plasmid carrying the cloned gene did not lead to an increase in autolysin production above the wild-type level, but it changed the colony morphology from smooth to rough. Among several autolysin-deficient mutations, lyt-151 was suppressed only by the high-copy-number plasmid carrying the cloned gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology
  • Bacillus subtilis / genetics*
  • Bacillus subtilis / physiology
  • Chromosomes, Bacterial
  • Cloning, Molecular
  • DNA, Bacterial / genetics
  • Flagella*
  • Genes, Bacterial*
  • N-Acetylmuramoyl-L-alanine Amidase / metabolism
  • Phenotype
  • Plasmids

Substances

  • DNA, Bacterial
  • N-Acetylmuramoyl-L-alanine Amidase