Isolation and Characterization of Specialized Lambda Transducing Bacteriophage Carrying the metBJF Methionine Gene Cluster

Abstract

Secondary attachment site lysogens of Δ att λ Δ ppc-argECBH strains of Escherichia coli with λ c I857 integrated into the bfe gene (88 min) were isolated. Of 20 such lysogens examined, 2 produce lysates with transducing phage containing the metBJF gene cluster (87 min). Reintroduction of the ppc-argECBH chromosome segment (which lies between the bfe and met genes) into these strains virtually abolishes the production of met transducing phage. All of the phage examined have lost essential genes from the left arm of the λ chromosome. Approximately 85% of the phage appear to have the same genetic composition, containing the metBJF gene cluster, but not the closely linked gene cytR , and having lost phage genes G and J . Analytical CsCl density gradient centrifugation of five representatives of this major class of phage shows four of them to have identical densities (lighter than λ), while the fifth cannot be resolved from λ. The four apparently identical phage were isolated from three separate lysates, which suggests the existence of preferred sites for illegitimate recombination on the bacterial and phage chromosomes. Three specialized transducing phage that carry cytR in addition to metB, metJ , and metF have also been studied. Each of these viruses has a different amount of phage deoxyribonucleic acid. Two of them have less deoxyribonucleic acid than λ, whereas the third has about the same amount. The metB, metF , and cytR genes of the transducing phage have been shown to function in vivo. The phage-borne metB and metF genes are subject to metJ -mediated repression.