biostudies-literatureUnknown77(1)Vilen HAdvances in DNA transposition technology have recently generated efficient tools for various types of functional genetic analyses. We demonstrate here the power of the bacteriophage Mu-derived in vitro DNA transposition system for modification and functional characterization of a complete bacterial virus genome. The linear double-stranded DNA genome of Escherichia coli bacteriophage PRD1 was studied by insertion mutagenesis with reporter mini-Mu transposons that were integrated in vitro into isolated genomic DNA. After introduction into bacterial cells by electroporation, recombinant transposon-containing virus clones were identified by autoradiography or visual blue-white screening employing alpha-complementation of E. coli beta-galactosidase. Additionally, a modified transposon with engiJournal of virology123-34https://www.ebi.ac.uk/biostudies/studies/S-EPMC140628biostudies-literatureA direct transposon insertion tool for modification and functional analysis of viral genomes.PMC140628Aalto JMKassinen ASavilahti HVilen HPaulin LfalseA direct transposon insertion tool for modification and functional analysis of viral genomes.Advances in DNA transposition technology have recently generated efficient tools for various types of functional genetic analyses. We demonstrate here the power of the bacteriophage Mu-derived in vitro DNA transposition system for modification and functional characterization of a complete bacterial virus genome. The linear double-stranded DNA genome of Escherichia coli bacteriophage PRD1 was studied by insertion mutagenesis with reporter mini-Mu transposons that were integrated in vitro into isolated genomic DNA. After introduction into bacterial cells by electroporation, recombinant transposon-containing virus clones were identified by autoradiography or visual blue-white screening employing alpha-complementation of E. coli beta-galactosidase. Additionally, a modified transposon with engi2003-01-01T00:00:00Z2003 Jan2025-04-04T14:48:35.737Z2019-03-27T00:17:30ZS-EPMC1406281247781710.1128/jvi.77.1.123-134.2003