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 π 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 mobiliz...

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Veröffentlicht in:	Gene 1992-09, Vol.118 (1), p.145-146
Hauptverfasser:	Penfold, Robert J., Pemberton, John M.
Format:	Artikel
Sprache:	eng
Schlagworte:	Bacterial Proteins - genetics Biological and medical sciences Biotechnology Chromosomes, Bacterial DNA Helicases DNA, Recombinant - genetics DNA-Binding Proteins Fundamental and applied biological sciences. Psychology Genetic engineering Genetic technics Genetic Vectors - genetics Methods. Procedures. Technologies mobilization Mutagenesis, Insertional - methods pir gene R6K replicon Recombinant DNA Replicon - genetics Rhodobacter sphaeroides Rhodobacter sphaeroides - genetics site-directed mutagenesis Trans-Activators Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Beschreibung
Zusammenfassung:	We have constructed an R6K-based suicide vector (pJP5603) that requires a trans supply of the pir-encoded π 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 α-complementation cloning. Vector functionality was demonstrated in Rhodobacter sphaeroides.
ISSN:	0378-1119 1879-0038
DOI:	10.1016/0378-1119(92)90263-O