A genetic system that reports transient activation of genes in Bacillus.
Site-specific recombination is a powerful tool for precise excision of DNA fragments. We used this characteristic to construct a genetic system to report the transient activation of a promoter by promoting the stable acquisition of an antibiotic resistance marker by the bacterium. The system is composed of two compatible plasmid derivatives from Gram-positive bacteria. One of the plasmids allows the insertion of promoters upstream from tnpI, which encodes the site-specific recombinase of Tn4430. The second plasmid carries two selectable resistance genes: one is flanked by two site-specific recombination sequences and is lost following recombination; in contrast, the other resistance gene becomes functional after the site-specific recombination event. By inserting conditionally controlled promoters (the xylose-inducible xylA promoter or the plcA promoter whose expression is dependent on the growth medium) upstream of tnpI, we demonstrated that our genetic system responds to signals inducing transcription by conferring a new resistance phenotype to the host bacteria. Thus, this system can be used to identify genes which are transiently or conditionally expressed.
Salamitou
S
S
Unité de Biochimie Microbienne, Institut Pasteur, Paris, France. mitou@pasteur.fr
Agaisse
H
H
Lereclus
D
D
D019677:Artificial Gene Fusion; D001407:Bacillus / Q000235:genetics*; D001409:Bacillus cereus / Q000201:enzymology / Q000235:genetics; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001413:Bacillus thuringiensis / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D004251:DNA Transposable Elements; D004254:DNA Nucleotidyltransferases; C069598:tnpI recombinase