USE OF GENETIC-RECOMBINATION AS A REPORTER OF GENE-EXPRESSION

Citation
A. Camilli et al., USE OF GENETIC-RECOMBINATION AS A REPORTER OF GENE-EXPRESSION, Proceedings of the National Academy of Sciences of the United Statesof America, 91(7), 1994, pp. 2634-2638
Citations number
23
Categorie Soggetti
Multidisciplinary Sciences
ISSN journal
00278424
Volume
91
Issue
7
Year of publication
1994
Pages
2634 - 2638
Database
ISI
SICI code
0027-8424(1994)91:7<2634:UOGAAR>2.0.ZU;2-9
Abstract
An understanding of the patterns of gene expression in response to spe cific environmental signals can yield insight into a variety of comple x biological systems such as microbial-host interactions, developmenta l cycles, cellular differentiation, ontogeny, etc. To extend the utili ty of the reporter gene fusion approach to such studies, we have const ructed a gene expression reporter cassette that permits the generation of transcriptional fusions to tnpR encoding resolvase, a site-specifi c recombinase of the transposable element gammadelta. 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 reso lvase functions. The loss of tetracycline resistance in descendant bac teria serves as a permanent and heritable marker of prior gene express ion. 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 inducin g environment facilitating the study of gene expression in complex env ironments such as animal tissues.