An improved suicide vector for construction of chromosomal insertion mutations in bacteria.
We have constructed an R6K-based suicide vector (pJP5603) that requires a trans supply of the pir-encoded pi protein of plasmid R6K for replication. Therefore, efficient plasmid suicide results upon transfer to bacteria not harbouring pir. The 3.1-kb vector encodes kanamycin resistance and is mobilizable. When used in conjunction with a JM109 strain carrying pir, it has nine unique restriction sites available for alpha-complementation cloning. Vector functionality was demonstrated in Rhodobacter sphaeroides.
Penfold
R J
RJ
Department of Microbiology, University of Queensland, St. Lucia, Australia.
Pemberton
J M
JM
D001426:Bacterial Proteins / Q000235:genetics*; D002876:Chromosomes, Bacterial; D004265:DNA Helicases; D004274:DNA, Recombinant / Q000235:genetics; D004268:DNA-Binding Proteins; D005822:Genetic Vectors / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods*; D012093:Replicon / Q000235:genetics; D012242:Rhodobacter sphaeroides / Q000235:genetics*; D015534:Trans-Activators
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D004265:DNA Helicases