Ooi Lezanne, Wood Ian C
Institute of Membrane and Systems Biology, University of Leeds, UK.
Methods Mol Biol. 2008;491:3-17. doi: 10.1007/978-1-59745-526-8_1.
A comprehensive understanding of regulatory protein interactions with their target genes is fundamental to determining transcriptional networks and identifying important events in the regulation of gene expression. Here we describe how transcriptional regulatory regions are to be identified using luciferase assays (including the transfection of cells by Amaxa and lipid-based reagents) and how protein-DNA interactions are to be characterised by chromatin immunoprecipitation (ChIP) coupled with quantitative PCR. Together these techniques provide a powerful combination for investigating potassium channel gene regulation.
全面了解调节蛋白与其靶基因的相互作用是确定转录网络和识别基因表达调控中重要事件的基础。在此，我们描述了如何使用荧光素酶测定法（包括使用Amaxa和基于脂质的试剂转染细胞）来鉴定转录调控区域，以及如何通过染色质免疫沉淀（ChIP）结合定量PCR来表征蛋白质 - DNA相互作用。这些技术共同为研究钾通道基因调控提供了强大的组合。
