Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu

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Abstract

A general procedure is described for transposing the lac genes to selected locations on the Escherichia coli chromosome. These transpositions were designed so that the lac genes could be fused to nearby promoters. In particular, the lac genes were fused to the promoters for the araBAD, araC and leu genes. In these fusions the lac genes are regulated by the controls of the genes to which they are fused. These fusions are therefore useful in discovering new types of regulation of gene expression, as was found in the case of the araC gene. λ transducing phage carrying the fusion as well as nearby genes can easily be isolated. Some of these fusions may result in the formation of hybrid proteins.

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    This work was supported by a National Institutes of Health pre-doctoral training grant and American Cancer Society grant VC-13 to J. Beckwith.

    Abbreviations used (Taylor & Trotter, 1972): lacZ and lacY are the only lac needed to be expressed for normal growth (Lac+ phenotype) on lactose. araC and lacI are control genes for the araOIBAD and lacPOZYA operons, leu and trpEDCBA are operons for genes involved in the biosynthesis of leucine and tryptophan. araOIBAD and lacPOZYA are operons for genes involved in the catabolism of arabinose and lactose.

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