An improved tetracycline-inducible expression vector for Staphylococcus aureus.
The tetracycline-inducible expression vector pALC2073 allowed high level expression of the cloned sasG gene but repression by uninduced cells was leaky. The -10 box of the tetR promoter was mutated to the Bacillus subtitlis consensus, which resulted in complete repression of SasG protein expression. Anhydrotetracycline at 1.28 microg ml(-1) gave the same high level of induction that was obtained with pALC2073sasG using 160 ng ml(-1) tetracycline, the highest concentration that could be used without inhibiting bacterial growth. This variant of pALC2073 thus offers almost complete repression when uninduced and high levels of expression when induced.
Corrigan
Rebecca M
RM
Department of Microbiology, Moyne Institute of Preventive Medicine, Trinity College, Dublin 2, Ireland.
Foster
Timothy J
TJ
D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005822:Genetic Vectors / Q000187:drug effects; D008565:Membrane Proteins / Q000235:genetics; D009154:Mutation / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects / Q000235:genetics; D013211:Staphylococcus aureus / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology*
D001426:Bacterial Proteins; D008565:Membrane Proteins; C524374:SasG protein, Staphylococcus aureus; D013752:Tetracycline