From an electrophoretic mobility shift assay to isolated transcription factors: a fast genomic-proteomic approach.

ABSTRACT:

Background

Hypocrea jecorina (anamorph Trichoderma reesei) is a filamentous ascomycete of industrial importance due to its hydrolases (e.g., xylanases and cellulases). The regulation of gene expression can influence the composition of the hydrolase cocktail, and thus, transcription factors are a major target of current research. Here, we design an approach for identifying a repressor of a xylanase-encoding gene.

Results

We used streptavidin affinity chromatography to isolate the Xylanase promoter-binding protein 1 (Xpp1). The optimal conditions and templates for the chromatography step were chosen according to the results of an electrophoretic mobility shift assay performed under repressing conditions, which yielded a DNA-protein complex specific to the AGAA-box (the previou

SUBMITTER: Mach-Aigner AR 

PROVIDER: S-EPMC3012607 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature