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

被引:11
作者
McCutcheon, Susan C. [1 ]
Jones, Ken [2 ]
Cumming, Sarah A. [3 ]
Kemp, Richard [1 ]
Ireland-Zecchini, Heather [1 ]
Saunders, John C. [4 ]
Houghton, Carol A. [1 ]
Howard, Louise A. [1 ]
Winton, Douglas J. [1 ]
机构
[1] Canc Res UK Cambridge Res Inst, Li Ka Shing Ctr, Cambridge CB2 0RE, England
[2] Univ Cambridge, Wellcome Trust Ctr Stem Cell Res, Cambridge CB2 1QR, England
[3] Univ Glasgow, Div Virol, Glasgow G11 5JR, Lanark, Scotland
[4] Babraham Inst, Gene Targeting Facil, Cambridge CB2 4AT, England
基金
英国生物技术与生命科学研究理事会;
关键词
ESCHERICHIA-COLI; MAMMALIAN-CELLS; GENE-EXPRESSION; CRE RECOMBINASE; STEM-CELLS; MARKER; LACZ;
D O I
10.1186/1741-7007-8-89
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 beta-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 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. Conclusions: SYNbglA will have general applicability to developmental and molecular studies in vitro and in vivo.
引用
收藏
页数:8
相关论文
共 27 条
[1]   IMPROVED IN-SITU BETA-GALACTOSIDASE STAINING FOR HISTOLOGICAL ANALYSIS OF TRANSGENIC MICE [J].
AGUZZI, A ;
THEURING, F .
HISTOCHEMISTRY, 1994, 102 (06) :477-481
[2]   An improved β-galactosidase reporter gene [J].
Anson, DS ;
Limberis, M .
JOURNAL OF BIOTECHNOLOGY, 2004, 108 (01) :17-30
[3]  
Arnone MI, 1997, DEVELOPMENT, V124, P4649
[4]  
Arnone MI, 2004, METHOD CELL BIOL, V74, P621
[5]   Transgenic mice expressing a dual, CRE-inducible reporter for the analysis of axon guidance and synaptogenesis [J].
Badaloni, Aurora ;
Bonanomi, Dario ;
Albieri, Ilaria ;
Givogri, Irene ;
Bongarzone, Ernesto ;
Valtorta, Flavia ;
Giacomo Consalez, G. .
GENESIS, 2007, 45 (06) :405-412
[6]   Identification of stem cells in small intestine and colon by marker gene Lgr5 [J].
Barker, Nick ;
van Es, Johan H. ;
Kuipers, Jeroen ;
Kujala, Pekka ;
van den Born, Maaike ;
Cozijnsen, Miranda ;
Haegebarth, Andrea ;
Korving, Jeroen ;
Begthel, Harry ;
Peters, Peter J. ;
Clevers, Hans .
NATURE, 2007, 449 (7165) :1003-U1
[7]   Talking about a revolution: The impact of site-specific recombinases on genetic analyses in mice [J].
Branda, CS ;
Dymecki, SM .
DEVELOPMENTAL CELL, 2004, 6 (01) :7-28
[8]   ECDYSTEROID-DEPENDENT REGULATION OF GENES IN MAMMALIAN-CELLS BY A DROSOPHILA ECDYSONE RECEPTOR AND CHIMERIC TRANSACTIVATORS [J].
CHRISTOPHERSON, KS ;
MARK, MR ;
BAJAJ, V ;
GODOWSKI, PJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (14) :6314-6318
[9]   A single type of progenitor cell maintains normal epidermis [J].
Clayton, Elizabeth ;
Doupe, David P. ;
Klein, Allon M. ;
Winton, Douglas J. ;
Simons, Benjamin D. ;
Jones, Philip H. .
NATURE, 2007, 446 (7132) :185-189
[10]   REPORTER GENES IN TRANSGENIC MICE [J].
CUI, CQ ;
WANI, MA ;
WIGHT, D ;
KOPCHICK, J ;
STAMBROOK, PJ .
TRANSGENIC RESEARCH, 1994, 3 (03) :182-194