Kamakaka R T, Kadonaga J T
Department of Biology, University of California, San Diego, La Jolla 92093.
Methods Cell Biol. 1994;44:225-35. doi: 10.1016/s0091-679x(08)60916-4.
We describe the preparation and use of a nuclear extract derived from Drosophila embryos that is highly active for transcription in vitro by RNA polymerase This extract, which is termed the soluble nuclear fraction (SNF), can support multiple rounds of transcription and generate about 0.45 transcripts per template per 30 min. Furthermore, the SNF is deficient in nonspecific DNA binding factors that inhibit transcription, such as histone H1, and can be used for the analysis of transcriptional regulation by promoter- and enhancer-binding factors with either naked DNA or chromatin templates.
我们描述了一种从果蝇胚胎中提取的核提取物的制备方法及其用途，该提取物对RNA聚合酶的体外转录具有高度活性。这种提取物被称为可溶性核组分（SNF），它能够支持多轮转录，并且每30分钟每个模板可产生约0.45个转录本。此外，SNF缺乏抑制转录的非特异性DNA结合因子，如组蛋白H1，可用于通过启动子和增强子结合因子对裸露DNA或染色质模板进行转录调控分析。
