Structural and functional analysis of the gene promoter-5

Copyright information:Taken from "Structural and functional analysis of the gene promoter"http://www.biomedcentral.com/1471-2199/8/82BMC Molecular Biology 2007;8():82-82.Published online 20 Sep 2007PMCID:PMC2064931. and its putative cis-acting consensus sequences (boxes). , schematic representation of the relevant features of the plasmids used for promoter activity assays. , 5'-flanking fragments of , containing -428 (pRab428), -415 (pRab415) (without the URE1-like sequence), and -369 (pRab369) nt from the first transcription initiation site., CAT activities obtained from trophozoites transfected with constructions showed at left. Activities are relative to that obtained with promoter gene, used as positive control. Each bar corresponds to the average of CAT activities ± S.D. representative of three independent experiments performed by duplicate. EMSA using the P-labeled sequences URE1 from (URE1 ) or from (URE ) gene promoters as probes and 30 μg of trophozoite nuclear extracts (NE). Lanes 1–5, EMSA using the URE1 sequence from as a probe. Lane 1, free probe. Lane 2, no competitor. Lane 3, specific competitor (SC) (150-fold excess of the same cold fragment). Lane 4, URE1 from the gene promoter used as competitor (URE1) (150-fold excess). Lane 5, unspecific competitor (UC) (150-fold excess of a mingled oligonucleotide with similar composition). Lanes 6–8, EMSA using the URE1 sequence from as a probe. Lane 6, no competitor. Lane 7, unspecific competitor (UC) (150-fold excess of a mingled oligonucleotide with similar composition). Lane 8, free probe. Arrowheads indicate the two DNA-protein complexes formed by probe-nuclear extracts interaction.