Gel mobility shift scanning of pectin-inducible promoter from Penicillium griseoroseum reveals the involvement of a CCAAT element in the expression of a polygalacturonase gene

Genetics and Molecular Biology

Andréa De O.b. Ribon²  João Batista Ribeiro¹  Daniel B. Gonçalves¹  Marisa V. De Queiroz¹  Elza F. De Araújo¹ 

[1] ,Universidade Federal de Viçosa Departamento de Bioquímica e Biologia Molecular Viçosa MG ,Brazil

关键词: Penicillium griseoroseum;    polygalacturonase;    5' upstream regulatory sequence;    electrophoretic mobility shift assay;   

DOI  :  10.1590/S1415-47572009005000021

来源: SciELO

【 摘 要 】

Previous reports have described pgg2, a polygalacturonase-encoding gene of Penicillium griseoroseum, as an attractive model for transcriptional regulation studies, due to its high expression throughout several in vitro growth conditions, even in the presence of non-inducing sugars such as sucrose. A search for regulatory motifs in the 5' upstream regulatory sequence of pgg2 identified a putative CCAAT box that could justify this expression profile. This element, located 270 bp upstream of the translational start codon, was tested as binding target for regulatory proteins. Analysis of a 170 bp promoter fragment by electrophoretic mobility shift assay (EMSA) with nuclear extracts prepared from mycelia grown in pectin-containing culture medium revealed a high mobility complex that was subsequently confirmed by analyzing it with a double-stranded oligonucleotide spanning the CCAAT motif. A substitution in the core sequence for GTAGG partially abolished the formation of specific complexes, showing the involvement of the CCAAT box in the regulation of the polygalacturonase gene studied.

【 授权许可】

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