title: A modified chromatin-immunoprecipitation protocol for silkmoth ovarian follicular cells reveals C/EBP and GATA binding modes on an early chorion gene promoter.
abstract: Standard Chromatin immunoprecipitation protocols have been designed to suit studies performed on cell line cultures or yeast cells growing in liquid cultures. In these cases cross-linking/fixation takes place directly in the growing medium of the cells by the addition of a general fixation reagent. When applied on whole isolated silkmoth follicles, this procedure results in poor release of follicular cells from the basal membrane and lower yield of cross-linked chromatin. We present a modification to the standard protocol, where detachment of follicular cells from the basal membrane of the egg and nuclei isolation precedes formaldehyde-mediated cross-linking. We also discuss application of the modified method for the identification of distinct BmC/EBP and BmGATAbeta binding modes on a chorion gene promoter from the Er1.A/B early gene pair.
authors: Department of Biochemistry and Molecular Biology, Faculty of Biology, National and Kapodistrian University of Athens, 15701 Athens, Greece.
authors: Argyris
authors: A
authors: Papantonis
authors: Rena
authors: R
authors: Lecanidou
mesh_terms: D000818:Animals; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002823:Chorion / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D005260:Female; D050980:GATA Transcription Factors / Q000378:metabolism*; D006080:Ovarian Follicle / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013997:Time Factors
chemical_list: D022762:CCAAT-Enhancer-Binding Proteins; D050980:GATA Transcription Factors