ChIP-nexus analysis of fission yeast transcription factors Cbf11, Cbf11DBM, Cbf12, Cbf12DBM, Mga2 and Fkh2

Genome-wide identification of transcription factor (TF) binding sites in the genome of the fission yeast Schizosaccharomyces pombe. The ChIP-nexus method was used. TFs included were: Cbf11-TAP and Cbf12-TAP (and their DBM mutants with impaired DNA binding), TAP-Mga2, and Fkh2-TAP (as an irrelevant control TF). IPs from an untagged WT strain were also analyzed. Cbf11-related IPs were performed from exponential cultures, while Cbf12-related IPs were performed from stationary cultures. YES complex medium was used for all cultivations.

本研究对裂殖酵母（Schizosaccharomyces pombe）基因组中的转录因子（Transcription Factor，TF）结合位点开展了全基因组范围鉴定。实验采用ChIP-nexus技术。本次研究选用的转录因子包括：Cbf11-TAP与Cbf12-TAP（及其DNA结合功能受损的DBM突变体）、TAP-Mga2，以及Fkh2-TAP（作为无关对照转录因子）。此外，本研究还对未标记野生型（Wild Type，WT）菌株的免疫沉淀（Immunoprecipitation，IP）样本进行了分析。其中，针对Cbf11的免疫沉淀实验于指数生长期培养物中开展，而针对Cbf12的免疫沉淀实验则于稳定期培养物中进行。所有培养均采用YES复合培养基。