ChIP-Seq of constitutively expressed Ndt80-Myc in K. lactis

The purpose of this experiment was to identify the genes bound by Ndt80 in K. lactis when constitutively expressed. Ndt80 was tagged with c-myc, the native Ndt80 promoter was replaced with the K. lactis Gal1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown overnight in SRaffinose until log-phase, then in SRaffinose + 1.7% Galactose for 5 hours. Each experiment was repeated twice and sequenced on an Illumina HiSeq 4000. K.lactis Ndt80-myc vs. untagged control, with two technical replicates for each

本实验旨在鉴定组成型表达状态下，乳酸克鲁维酵母（K. lactis）中被Ndt80结合的基因。具体实验流程如下：将Ndt80以c-myc标签进行标记，将内源Ndt80启动子替换为乳酸克鲁维酵母Gal1启动子，随后使用c-myc抗体进行免疫沉淀。细胞先在SRaffinose培养基中过夜培养至对数生长期，再转移至添加1.7%半乳糖的SRaffinose培养基中培养5小时。本实验共重复两次，所有样本均在Illumina HiSeq 4000测序平台完成测序；实验组为K.lactis Ndt80-myc，对照组为未标记样本，每组均设置两个技术重复。