Characterization of a heat resistant beta-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 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.
McCutcheon
Susan C
SC
Cancer Research UK Cambridge Research Institute, Li Ka Shing Centre, UK.
Jones
Ken
K
Cumming
Sarah A
SA
Kemp
Richard
R
Ireland-Zecchini
Heather
H
Saunders
John C
JC
Houghton
Carol A
CA
Howard
Louise A
LA
Winton
Douglas J
DJ
D000818:Animals; D002460:Cell Line; D004795:Enzyme Stability; D015870:Gene Expression; D017930:Genes, Reporter; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D041681:NIH 3T3 Cells; D046968:Thermoanaerobacter / Q000201:enzymology*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics*
D001616:beta-Galactosidase