New ccdB positive-selection cloning vectors with kanamycin or chloramphenicol selectable markers.
Recently, we described pKIL18/19 positive-selection cloning vectors which rely on the inactivation of the cytotoxic ccdB gene [Bernard et al., Gene 148 (1994) 71-74]. They are new tools for simplifying gene cloning/sequencing procedures, as only recombinant DNA allow the formation of viable colonies. To enhance positive-selection capabilities, putative translational reinitiation codons located within the pKIL18/19 ccdB were modified by site-directed mutagenesis. New pKIL-derived vectors with kanamycin (KmR) or chloramphenicol (CmR) resistance-encoding genes were also constructed. In addition, a new host derived from the DH2 strain was developed.
Bernard
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Département de Biologie Moléculaire, Université Libre de Bruxelles, Rhode-Saint-Genèse, Belgium.
D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D002702:Chloramphenicol Resistance; D003001:Cloning, Molecular / Q000379:methods*; D005819:Genetic Markers; D005822:Genetic Vectors; D007613:Kanamycin Resistance; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012641:Selection, Genetic
D001426:Bacterial Proteins; D001427:Bacterial Toxins; C068356:CcdB protein, Plasmid F; D005819:Genetic Markers