A specialized transducing phage, lambda psrlA, for the sorbitol phosphotransferase of Escherichia coli K12.
A specialized transducing phage for the srlA gene, specifying the sorbitol-specific Enzyme II of the phosphoenolpyruvate:sugar phosphotransferase system, was constructed and its DNA was analysed by restriction endonuclease digestion. Phage construction involved four steps: (1) integration of lambda into the srlA gene; (2) selection of phage carrying (a) the left and (b) the right end of the srlA gene by means independent of the function of the new DNA acquired; (3) reconstitution of the srlA gene in a dilysogen of these two phage; and (4) the excision, using the heteroimmune lambdoid phage 21, of a plaque-forming srlA+ phage from the dilysogenic chromosome. Comparison of the DNA restriction digests of the transducing phage with those of its parents and of wild-type lambda revealed fragments consisting partly of lambda and partly of Escherichia coli DNA. The junction points in the intermediate phage define a site that must lie within the reconstituted gene of the final phage. This technique should be of general application in relating genes, cloned by our method, to DNA sequences.
D010582:Bacteriophage lambda / Q000235:genetics*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D005814:Genes, Viral; D008242:Lysogeny; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics*; D014161:Transduction, Genetic
D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C034466:phosphoenolpyruvate-sorbitol phosphotransferase; D004262:DNA Restriction Enzymes