Use of genetic recombination as a reporter of gene expression.
An understanding of the patterns of gene expression in response to specific environmental signals can yield insight into a variety of complex biological systems such as microbial-host interactions, developmental cycles, cellular differentiation, ontogeny, etc. To extend the utility of the reporter gene fusion approach to such studies, we have constructed a gene expression reporter cassette that permits the generation of transcriptional fusions to tnpR encoding resolvase, a site-specific recombinase of the transposable element gamma delta. Induction of the transcriptional fusions results in production of resolvase, which in turn, catalyzes excision of a linked tetracycline-resistance reporter gene flanked by direct repeats of res, the DNA sequences at which resolvase functions. The loss of tetracycline resistance in descendant bacteria serves as a permanent and heritable marker of prior gene expression. This gene fusion approach will allow us to assay the induction of gene expression in as few as one cell. Additionally, gene expression can be assayed at a later time and/or different place from the inducing environment facilitating the study of gene expression in complex environments such as animal tissues.
Camilli
A
A
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115.
Beattie
D T
DT
Mekalanos
J J
JJ
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D017930:Genes, Reporter; D051379:Mice; D008815:Mice, Inbred Strains; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009713:Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D013753:Tetracycline Resistance / Q000235:genetics; D019895:Transposases; D014734:Vibrio cholerae / Q000235:genetics* / Q000472:pathogenicity; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C074789:irgA protein, Vibrio cholerae; D009713:Nucleotidyltransferases; D019895:Transposases; D000469:Alkaline Phosphatase