Ontology type: schema:ScholarlyArticle
1980-11
AUTHORSDavid W. Ow, Frederick M. Ausubel
ABSTRACTWe 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. More... »
PAGES165-175
http://scigraph.springernature.com/pub.10.1007/bf00267366
DOIhttp://dx.doi.org/10.1007/bf00267366
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PUBMEDhttps://www.ncbi.nlm.nih.gov/pubmed/6255293
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