PMID:4607764

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Citation

Silhavy, TJ and Boos, W (1974) Selection procedure for mutants defective in the beta-methylgalactoside transport system of Escherichia coli utilizing the compound 2R-glyceryl-beta-D-galactopyranoside. J. Bacteriol. 120:424-32

Abstract

A procedure has been devised that allows selection of mutants defective in the beta-methylgalactoside transport system (mgl) of Escherichia coli. This procedure utilizes the compound 2R-glyceryl-beta-d-galactopyranoside (glycerylgalactoside), which is known to be transported by only two transport system in E. coli, namely, the lactose and the beta-methylgalactoside transport systems. Mutants lacking glycerol-3-phosphate dehydrogenase (glpD) are sensitive to glycerol. Similarly, mutants lacking uridine diphosphate-galactose-4-epimerase (galE) are sensitive to galactose. Glycerylgalactoside is an inducer of the lactose operon and also a substrate for beta-galactosidase. Thus, a mgl(+)glpD galE lacY strain will not grow in the presence of glycerylgalactoside owing to accumulated glycerol-3-phosphate, galactose-1-phosphate, and uridine diphosphate-galactose. We have constructed such a strain and shown that mgl mutants can be obtained by selecting for those that grow in the presence of glycerylgalactoside.

Links

PubMed PMC245779

Keywords

Autoradiography; Biological Transport, Active; Carbohydrate Epimerases/biosynthesis; Carbon Radioisotopes; Chromatography, Paper; Escherichia coli/enzymology; Escherichia coli/isolation & purification; Escherichia coli/metabolism; Galactose/metabolism; Galactosidases/biosynthesis; Glycerol/metabolism; Glycerolphosphate Dehydrogenase/biosynthesis; Glycosides/metabolism; Kinetics; Methylgalactosides/metabolism; Methylglycosides/metabolism; Mutagens; Mutation; Nitrosoguanidines; Succinates/metabolism; Transduction, Genetic

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