A novel plasmid vector that uses the glucose kinase gene (glkA) for the positive selection of stable gene disruptants in Streptomyces.
We describe an Escherichia coli plasmid, pIJ2581, that can be used for the efficient construction of stable gene disruptants and of gene deletions in Streptomyces. Integration of pIJ2581 derivatives carrying chromosomal sequences is achieved by selecting for plasmid-encoded thiostrepton resistance, while plasmid excision is secured by counter-selection of the pIJ2581 glkA gene, which confers sensitivity to 2-deoxyglucose.
van Wezel
G P
GP
John Innes Centre, Norwich Research Park, Colney, UK.
Bibb
M J
MJ
D001426:Bacterial Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005822:Genetic Vectors; D005941:Glucokinase / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D015183:Restriction Mapping; D017384:Sequence Deletion / Q000235:genetics; D013302:Streptomyces / Q000235:genetics*
D001426:Bacterial Proteins; C096446:GlkA protein, Bacteria; D005941:Glucokinase