Specific repression of the luciferase reporter gene by GBD-MTase fusion constructs

Copyright information:Taken from "Chimeric DNA methyltransferases target DNA methylation to specific DNA sequences and repress expression of target genes"Nucleic Acids Research 2006;35(1):100-112.Published online 06 Dec 2006PMCID:PMC1761428.© 2006 The Author(s) () GBD-3a, GBD-3b and GBD expression constructs were co-transfected with firefly luciferase reporter plasmids 5×UAS-TK-Luc or TK-Luc into HEK293T cells and the reporter gene activity determined. () Co-transfection of the 5×UAS-TK-Luc reporter plasmid with GBD, GBD-3a, mutGBD-3a and GBD-3a-ANV/E74A expression vectors into HEK293T cells. () Co-transfection of the 5×UAS-TK-Luc reporter plasmid with GBD, GBD-3b, mutGBD-3b and GBD-3b-ANV expression vectors into HEK293T cells. () Co-transfection of the UAS-TK-Luc reporter plasmid with GBD, GBD-3a, GBD-3a-ANV/E74A, GBD-3b and GBD-3b-ANV into HEK293T cells. () Co-transfection of UAS-ras-Luc and ras-Luc reporter plasmids with the GBD, GBD-3a, GBD-3a-ANV/E74A, GBD-3b and GBD-3b-ANV into HEK293T cells. In each case the error bars give the standard deviations of at least three independent experiments. () Competition experiment of GBD-3a and GBD. To investigate the targeting function of the Zinc finger, GBD-3a or GBD-3b were co-transfected with the 5×UAS-TK-Luc reporter plasmid construct together with increasing amounts of a construct that expresses GBD. The total amount of DNA for transfection was normalized by empty vector. The numbers below the axis indicate the amount of DNA of each construct (ng/well in 24-well plate) used in each transfection.