Bacteriophage P1 pac sites inserted into the chromosome greatly increase packaging and transduction of Escherichia coli genomic DNA

摘要: The Escherichia coli bacteriophage P1 packages host chromosome separately from phage DNA, and transfers it to recipient cells at low frequency in a process called generalized transduction. Phage genomes are packaged concatemers beginning specific site, pac. To increase transduction rate, we have inserted pac into the up five equally spaced positions; least this many fully tolerated absence of infection. A single chromosomal greatly increases downstream markers without decreasing yields; 3.5 × as much total DNA is packaged. Additional insertions decrease yield by > 90% also synthesis, although less dramatically. Packaging near each site is, same time, increased greater than 10 fold. Transduction an can be over 1000-fold, potentially allowing identification such transductants screening.