Cloning of mglB, the structural gene for the galactose-binding protein of Salmonella typhimurium and Escherichia coli.
From libraries of EcoRI fragments of Salmonella typhimurium and Escherichia coli DNA in lambda gt7, phages could be isolated that carry mglB, the structural gene of the galactose-binding protein as well as other mgl genes. Lysogenization of an E. coli mutant carrying a defective galactose-binding protein with lambda gt7 mglB (Salmonella) restores full galactose transport and galactose chemotaxis. Both the E. coli mutant protein as well as the wild-type Salmonella galactose-binding protein are synthesized in this strain. The EcoR1 fragments of both organisms carrying the mgl genes were 6 Kb long. They were subcloned into the multicopy plasmid pACYC184. The hybrid plasmid containing the Salmonella mgl DNA gives rise to the synthesis of large amounts of galactose-binding protein in the periplasm of E. coli. The protein can be precipitated by antibodies against the E. coli binding protein and is identical to the fully processed protein isolated from Salmonella typhimurium LT2. In vitro protein synthesis (Zubay-system) with either lambda gt7 mgl phages as well as the hybrid plasmid as DNA matrix produces the galactose-binding protein mainly in precursor form that is precipitable by specific antibodies.
D010582:Bacteriophage lambda / Q000235:genetics; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins / Q000235:genetics*; D002633:Chemotaxis; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D005796:Genes; D005838:Genotype; D007700:Kinetics; D009004:Monosaccharide Transport Proteins; D009154:Mutation; D033902:Periplasmic Binding Proteins; D012486:Salmonella typhimurium / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity
D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; C023550:galactose-binding protein; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI; D005690:Galactose