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Analysis of developmentally regulated chorion gene promoter architecture via electroporation of silk moth follicles
Author(s) -
Tsatsarounos S. P.,
Rodakis G. C.,
Lecanidou R.
Publication year - 2015
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/imb.12136
Subject(s) - biology , promoter , gene , electroporation , reporter gene , genetics , gene expression , repressor , transcription factor , regulation of gene expression , microbiology and biotechnology
In the silk moth B ombyx mori , chorion genes of the same developmental specificity are organized in divergently transcribed α / β gene pairs, sharing a common 5′ flanking promoter region. This bidirectional promoter contains a complete set of cis ‐elements responsible for developmentally accurate gene expression. In the present paper, based on the observation that B ombyx chorion gene promoters contain cis ‐elements for the same transcription factors without concrete evidence on which of them are essential, we address the question as to how promoter architecture (number, orientation and position of common factor binding sites) facilitates developmentally accurate chorion gene regulation. To this end, we constructed several mutated promoter regions of an early‐middle gene pair and cloned them upstream of a reporter gene to introduce these plasmid constructs into silk moth follicle epithelial cells via electroporation as an efficient and quick method for transient expression. This is the first time that an ex vivo method had been applied to test the impact of systematic cis ‐element mutations on a chorion gene promoter. Our results confirmed the importance of the HMGA factor and the role of the GATA factor as an early repressor, and led to a more detailed understanding of which C / EBP sites participate in the regulation of early‐middle chorion gene expression.