Characterization of a heat resistant [beta]-glucosidase as a new reporter in cells and mice

• Published in: BMC biology Vol. 8; pp. 89 - 177
• Main Authors: McCutcheon, Susan C, Jones, Ken, Cumming, Sarah A, Kemp, Richard, Ireland-Zecchini, Heather, Saunders, John C, Houghton, Carol A, Howard, Louise A, Winton, Douglas J
• Format: Journal Article
• Language: English
• Published: BioMed Central Ltd 22-06-2010
• Subjects: Biological research; Biology, Experimental; Escherichia coli; Gene expression; Genetic aspects; Histochemistry; Mammals; Methods; Physiological aspects
• ISSN: 1741-7007; 1741-7007
• DOI: 10.1186/1741-7007-8-89

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 [beta]-glucosidase. 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 [beta]-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. SYNbglA will have general applicability to developmental and molecular studies in vitro and in vivo.