Molecular cloning of a gene affecting the autolysin level and flagellation in Bacillus subtilis.
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.
Sekiguchi
J
J
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan.
Ezaki
B
B
Kodama
K
K
Akamatsu
T
T
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005407:Flagella; D005798:Genes, Bacterial; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000378:metabolism; D010641:Phenotype; D010957:Plasmids
D004269:DNA, Bacterial; D009238:N-Acetylmuramoyl-L-alanine Amidase