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

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-gluc...

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Autores principales: 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
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2909947/
https://www.ncbi.nlm.nih.gov/pubmed/20569471
http://dx.doi.org/10.1186/1741-7007-8-89
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author 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
author_facet 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
author_sort McCutcheon, Susan C
collection PubMed
description 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.
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spelling pubmed-29099472010-07-27 Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice 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 BMC Biol Methodology Article 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. BioMed Central 2010-06-22 /pmc/articles/PMC2909947/ /pubmed/20569471 http://dx.doi.org/10.1186/1741-7007-8-89 Text en Copyright ©2010 McCutcheon et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
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
Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
title Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
title_full Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
title_fullStr Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
title_full_unstemmed Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
title_short Characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
title_sort characterization of a heat resistant ß-glucosidase as a new reporter in cells and mice
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2909947/
https://www.ncbi.nlm.nih.gov/pubmed/20569471
http://dx.doi.org/10.1186/1741-7007-8-89
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