Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
| Autor: | Susan C McCutcheon, Heather Ireland-Zecchini, Sarah A. Cumming, John Saunders, Richard Kemp, Louise Howard, Carol Houghton, Kenneth D Jones, Douglas J. Winton |
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| Rok vydání: | 2010 |
| Předmět: |
Hot Temperature
Physiology Transgene Gene Expression Mice Transgenic Thermoanaerobacter Plant Science Biology General Biochemistry Genetics and Molecular Biology Cell Line Mice 03 medical and health sciences 0302 clinical medicine Genes Reporter Structural Biology In vivo Enzyme Stability Gene expression Animals Humans Gene silencing lcsh:QH301-705.5 Gene Ecology Evolution Behavior and Systematics 030304 developmental biology 0303 health sciences Reporter gene Agricultural and Biological Sciences(all) Biochemistry Genetics and Molecular Biology(all) Methodology Article Cell Biology beta-Galactosidase Molecular biology In vitro lcsh:Biology (General) 030220 oncology & carcinogenesis NIH 3T3 Cells Bioreporter General Agricultural and Biological Sciences Developmental Biology Biotechnology |
| Zdroj: | BMC Biology BMC Biology, Vol 8, Iss 1, p 89 (2010) |
| ISSN: | 1741-7007 |
| DOI: | 10.1186/1741-7007-8-89 |
| Popis: | Background Reporter genes are widely used in biology and only a limited number are available. We present a new reporter gene for the localization of mammalian cells and transgenic tissues based on detection of the bglA (SYNbglA) gene of Caldocellum saccharolyticum that encodes a thermophilic β-glucosidase. Results SYNbglA was generated by introducing codon substitutions to remove CpG motifs as these are associated with gene silencing in mammalian cells. SYNbglA expression can be localized in situ or detected quantitatively in colorimetric assays and can be co-localized with E. coli β-galactosidase. Further, we have generated a Cre-reporter mouse in which SYNbglA is expressed following recombination to demonstrate the general utility of SYNbglA for in vivo analyses. SYNbglA can be detected in tissue wholemounts and in frozen and wax embedded sections. Conclusions SYNbglA will have general applicability to developmental and molecular studies in vitro and in vivo. |
| Databáze: | OpenAIRE |
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