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Open Access Methodology article

Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice

Susan C McCutcheon1, Ken Jones2, Sarah A Cumming3, Richard Kemp1, Heather Ireland-Zecchini1, John C Saunders4, Carol A Houghton1, Louise A Howard1 and Douglas J Winton1*

Author Affiliations

1 Cancer Research UK Cambridge Research Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK

2 Wellcome Trust Centre for Stem Cell Research, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK

3 Division of Virology, University of Glasgow, Glasgow G11 5JR, UK

4 Gene Targeting Facility, Babraham Institute, Babraham, Cambridge CB2 4AT, UK

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BMC Biology 2010, 8:89  doi:10.1186/1741-7007-8-89

Published: 22 June 2010

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.