Abstract
Preparation of nuclear extracts is a critical step for biochemical identification of factors which function in the nuclei such as transcription factors. We have established a new method to prepare nuclear extracts from prepupae or pupae of Drosophila melanogaster. The method is simple and particularly useful for small-scale isolation from materials that are hard to get in large amounts such as specific tissues or animal bodies covered by pupal cases.
Keywords: DNA binding factor, Drosophila, Gel mobility shift assay, Metamorphosis, Nuclear extract
Full Text
The Full Text of this article is available as a PDF (264.9 KB).
References
- Biggin M.D., Tijan R. Transcription factors that activate the Ultrabithorax promoter in developmentally staged extracts. Cell. 1988;53:699–711. doi: 10.1016/0092-8674(88)90088-8. [DOI] [PubMed] [Google Scholar]
- Jowett T., White K. Isolation of nuclei from embryos. In: Ashburner M., editor. Drosophila A LABORATORY MANUAL. Cold Spring Harbor, New York, USA: Cold Spring Harbor Laboratory Press; 1989. pp. 242–243. [Google Scholar]
- Kageyama Y., Masuda S., Hirose S., Ueda H. Temporal regulation of the mid-prepupal gene FTZ-F1: DHR3 early late gene product is one of the plural positive regulators. Genes Cells. 1997;2:559–569. doi: 10.1046/j.1365-2443.1997.1460344.x. [DOI] [PubMed] [Google Scholar]
- Kawasaki H., Hirose S., Ueda H. βFTZ-F1 dependent and independent activation of Edg78Ea pupal cuticle geneduring the early metamorphic period in Drosophila melanogaster. Dev. Growth Differ. 2002;44:419–425. doi: 10.1046/j.1440-169X.2002.00655.x. [DOI] [PubMed] [Google Scholar]
- Murata T., Kageyama Y., Hirose S., Ueda H. Regulation of the EDG84A gene by FTZ-F1 during metamorphosis in Drosophila melanogaster. Mol. Cell Biol. 1996;16:6509–6515. doi: 10.1128/mcb.16.11.6509. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ueda H., Hirose S. Identification and purification of a Bombyx mori homologue of FTZ-F1. Nucl. Acids Res. 1990;18:7229–7234. doi: 10.1093/nar/18.24.7229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ueda H., Sonoda S., Brown J.L., Scott M.P., Wu C. A sequence-specific DNA-binding protein that activates fushi tarazu segmentation gene expression. Genes Dev. 1990;4:624–635. doi: 10.1101/gad.4.4.624. [DOI] [PubMed] [Google Scholar]