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

J. Sekiguchi, B. Ezaki, K. Kodama, T. Akamatsu

Research output: Contribution to journalArticle

30 Citations (Scopus)

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 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.

Original languageEnglish
Pages (from-to)1611-1621
Number of pages11
JournalJournal of general microbiology
Volume134
Issue number6
DOIs
Publication statusPublished - 1988

ASJC Scopus subject areas

  • Microbiology

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