Characterization of a heat resistant beta-glucosidase as a new reporter in cells and mice
McCutcheon, Susan C
Cumming, Sarah A
Saunders, John C
Houghton, Carol A
Howard, Louise A
Winton, Douglas James
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McCutcheon, S. C., Jones, K., Cumming, S. A., Kemp, R., Ireland-Zecchini, H., Saunders, J. C., Houghton, C. A., et al. (2010). Characterization of a heat resistant beta-glucosidase as a new reporter in cells and mice.
Abstract 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.
This record's URL: http://www.dspace.cam.ac.uk/handle/1810/237611
Rights Holder: McCutcheon et al.; licensee BioMed Central Ltd.