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Recombinant P4 bacteriophages propagate as viable lytic phages or as autonomous plasmids in Klebsiella pneumoniae

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Summary

We demonstrate the use of bacteriophage P4 as a molecular cloning vector in Klebsiella pneumoniae. A hybrid P4 phage, constructed in vitro, that contains a K. pneumoniae hisDG DNA fragment can be propagated either as a lytic viable specialized transducing phage or as an autonomous, self-replicating plasmid. Hybrid P4 genomes existing as plasmids can be readily converted into non-defective P4-hybrid phage particles by superinfection with helper phage P2. Infection of a K. pneumoniae hisD non-P2 lysogen with P4-hisD hybrid phage results in approximately 90% of the infected cells becoming stably transduced to HisD+. Because P4 interferes with P2 growth, high titre stocks of P4 hybrid phages are relatively free (≤10-6) of P2 contamination. The hisG gene product was detected in ultraviolet light irradiated host cells infected by the P4-hisDG hybrid phage. A mutant of P4 (P4sidl) that directs the packaging of P4 DNA into P2 sized capsids should permit the construction of hybrid phages carrying 26 kilobase inserts.

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Authors and Affiliations

  1. Cellular and Developmental Group, Department of Biology, Biological Laboratories, Harvard University, 02138, Cambridge, Massachusetts, USA

    David W. Ow & Frederick M. Ausubel

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  1. David W. Ow
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  2. Frederick M. Ausubel
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Communicated by A.J. Clark

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Ow, D.W., Ausubel, F.M. Recombinant P4 bacteriophages propagate as viable lytic phages or as autonomous plasmids in Klebsiella pneumoniae . Molec. Gen. Genet. 180, 165–175 (1980). https://doi.org/10.1007/BF00267366

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  • DOI: https://doi.org/10.1007/BF00267366

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