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J Bacteriol. 1972 October; 112(1): 40-47
Copyright © 1972 American Society for Microbiology. All Rights Reserved.

Isolation of Transducing Bacteriophages for the Histidine and Isoleucine-Valine Operons in Escherichia coli K-12

¹ Centro di Endocrinologia ed Oncologia Sperimentale del CNR, Istituto di Patologia Generale dell'Università, 80138 Naples, Italy, and Istituto Internazionale di Genetica e Biofisica, Via Marconi 10, Naples, Italy

ABSTRACT

In vitro studies have been of great value in elucidating the mechanism of the regulation of several bacterial operons. To obtain a deoxyribonucleic acid preparation enriched for the histidine (his) and for the isoleucine-valine (ilv) operons, we have isolated bacteriophages carrying the his and the ilv regions of the Escherichia coli chromosome. Transposition of the his operon to a site close to the att80 region of the E. coli chromosome has been carried out selecting for integration of a temperature-sensitive F'his⁺ in the tonB locus. This transposed strain has been lysogenized with 80i. Upon induction of the lysogen, His⁺ transductants have been isolated, which, on further induction give rise to HFT (high frequency of transduction) lysates. Preliminary characterization of the transducing phage is reported. The ilv operon, carried on an F' particle, has been fused to an episome carrying the att80 region. The fused episome has been lysogenized with 80i t68. Upon induction of the lysogen, Ilv⁺ transductants have been isolated which on further induction give rise to HFT lysates.

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