Enzymatic sorting of bacterial colonies on filter paper replicas: detection of labile activities.
To utilize autoradiographic colony-sorting techniques (C. R. H. Raetz, Proc. Natl. Acad. Sci. U.S.A. 72:2274-2278, 1975) for the isolation of mutants with unstable enzymes, we report a new desiccation-induced lysis method, compatible with low temperatures. Furthermore, a general, two-step protocol is presented for clonal detection of hydrolytic reactions. The advantages of these critical modifications are demonstrated with the membrane enzymes glycerol 3-phosphate acyltransferase and cytidine 5'-diphosphate-diglyceride hydrolase.
D000217:Acyltransferases / Q000378:metabolism*; D001345:Autoradiography; D001431:Bacteriological Techniques; D003567:Cytidine Diphosphate Diglycerides / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000378:metabolism*; D009154:Mutation; D011755:Pyrophosphatases / Q000378:metabolism*
D003567:Cytidine Diphosphate Diglycerides; D000217:Acyltransferases; D005992:Glycerol-3-Phosphate O-Acyltransferase; D011755:Pyrophosphatases; C019855:CDPdiacylglycerol pyrophosphatase