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Figure 5 | BMC Molecular Biology

Figure 5

From: Structural and functional analysis of the Entamoeba histolytica EhrabB gene promoter

Figure 5

Delimitation of the activating region of EhrabB transcription. (A) Schematic representation of the EhrabB promoter region analyzed and its putative cis-acting consensus sequences (boxes). Arrows indicate the transcription initiation sites. (B) Left, schematic representation of the relevant features of the plasmids used for promoter activity assays. EhrabB, 5'-flanking fragments of EhrabB containing from -428 to -257 and +96 nt from the first transcription initiation site (+96).Right, CAT activities obtained from trophozoites transfected with constructions showed at left. Activities are relative to that obtained with the actin 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. (C) EMSA using the 32P-labeled fragment from -428 to -369 of EhrabB promoter as a probe and 30 μg of nuclear extracts (NE) from trophozoites. Lane 1, free probe. Lane 2, no competitor. Lane 3, specific competitor (SC) (150-fold excess of the same cold fragment). Lane 4, unspecific competitor (UC) (350-fold excess of poly [d(I-C)]). Arrowheads indicate the two DNA-protein complexes formed by interaction of the probe with NE.

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