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The use of specialised transducing phages in the amplification of enzyme production

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Summary

Two types of λtrp phages have been used as model systems to investigate ways of optimising the expression of bacterial genes from transducing phage genomes.

Excellent yields of trp enzymes were achieved by infecting a trpR− host with Q − or Q − Q − S − derivatives of λtrpAM1, which expresses its trp genese exclusively from the trp promoter. The five trp geneproducts constituted more than 50% of the total soluble protein of infected cells under these conditions, and an even higher proportion of the protein synthesized after infection. In a trpR + host, phage DNA replication was easily able to override tryptophan-mediated repression by titration of the trp repressor protein. N − derivatives of λtrp phages carrying the trp promoter were equally productive, while having the advantage of being much simpler to construct and propagate.

λtrp phages lacking the trp promoter were used to investigate ways of optimising gene expression initiated at the phage promoter, PL. Though very powerful, the latter promoter is more difficult to harness then the trp promoter. Derepression of transcription from PL by the use of cro − mutations is accompanied by poor replication of transducing phage DNA. Attempts to circumvent this difficulty using virulent of cro, cll double mutants have not been successful. Nevertheless, cells infected with a λtrp phage expressing its trp genes exclusively from PL made up to 16 per cent of their protein as trp gene-products.

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  1. Anne Moir

    Present address: Department of Genetics, University of Birmingham, B15 2TT, Birningham

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  1. Department of Molecular Biology, University of Edinburgh, Edinburgh, UK

    Anne Moir & W. J. Brammar

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  1. Anne Moir
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Communicated by B.A. Bridges

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Moir, A., Brammar, W.J. The use of specialised transducing phages in the amplification of enzyme production. Molec. Gen. Genet. 149, 87–99 (1976). https://doi.org/10.1007/BF00275963

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