Procedure for the isolation of mutants of Bacillus subtilis with defective cytoplasmic membranes.
A procedure has been devised to isolate mutants of Bacillus subtilis with structurally defective membranes. The procedure used to screen for the mutants involved comparison of the stability of protoplasts of the mutant with those of the wild type in a medium of sufficient osmotic strength to stabilize wild-type protoplasts. Mutagenized cells were grown as clones on agar plates, and then replicated onto plates containing 0.5 m lactose, which is sufficient to stabilize wild-type protoplasts. The colonies on the lactose-containing plates were then treated with lysozyme to convert the cells to protoplasts. Colonies of wild-type protoplasts remained opaque; however, colonies of mutant protoplasts lysed and became clear. Twenty-nine osmotically fragile mutants were isolated in this manner; the membranes of several mutants were found to contain alterations in the composition of their proteins or lipids.
D001412:Bacillus subtilis / Q000032:analysis / Q000166:cytology / Q000254:growth & development / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000032:analysis; D001431:Bacteriological Techniques; D002247:Carbon Isotopes; D002462:Cell Membrane / Q000032:analysis; D002845:Chromatography; D002999:Clone Cells; D003470:Culture Media; D005069:Evaluation Studies as Topic; D005227:Fatty Acids / Q000032:analysis; D005821:Genetic Techniques; D008055:Lipids / Q000032:analysis; D009113:Muramidase; D009153:Mutagens; D009154:Mutation; D009604:Nitrosoguanidines; D009994:Osmolar Concentration; D010743:Phospholipids / Q000032:analysis; D010759:Phosphorus Isotopes; D011523:Protoplasts; D014316:Tritium
D001426:Bacterial Proteins; D002247:Carbon Isotopes; D003470:Culture Media; D005227:Fatty Acids; D008055:Lipids; D009153:Mutagens; D009604:Nitrosoguanidines; D010743:Phospholipids; D010759:Phosphorus Isotopes; D014316:Tritium; D009113:Muramidase