Genome-wide maps of LDL1pro::LDL1:GFP in 14 days old arabidopsis

We report the application of single-molecule-based sequencing technology for high-throughput profiling of H3K4 demethylase LDL1 in 14 days old arabidopsis. By obtaining sequence from chromatin immunoprecipitated DNA, we generated genome-wide LDL1-binding maps of 14 days old arabidopsis. To reveal bound genes by LDL1, chimeric protein LDL1-GFP was expressed under LDL1 promoter in ldl1 (LDL1pro:LDL1:GFP/ ldl1). ChIP was performed using anti-GFP antibody (ab290; ABCAM), and ChIP DNA were analyzed by Illumina HiSeq 2500. Overall design: Chromatin immunoprecipitation was performed for LDL1pro::LDL1:GFP-expressing Arabidopsis. ChIP DNA was analyzed by Illumina HiSeq 2500.本研究报道了基于单分子测序技术（single-molecule-based sequencing technology）在14日龄拟南芥（Arabidopsis）中对组蛋白H3K4去甲基化酶LDL1开展高通量谱型分析的应用。通过对染色质免疫沉淀DNA（chromatin immunoprecipitated DNA, ChIP DNA）进行测序，我们获得了14日龄拟南芥全基因组范围内的LDL1结合图谱。为鉴定LDL1的靶结合基因，我们在ldl1突变体背景下，以LDL1自身启动子驱动嵌合蛋白（chimeric protein）LDL1-GFP的表达，构建了基因型为LDL1pro:LDL1:GFP/ ldl1的转基因材料。本实验使用抗GFP抗体（ab290；ABCAM公司）完成染色质免疫沉淀实验，并通过Illumina HiSeq 2500平台对ChIP DNA进行测序分析。整体实验设计：针对表达LDL1pro::LDL1:GFP的拟南芥材料开展染色质免疫沉淀实验，随后通过Illumina HiSeq 2500平台对ChIP DNA进行测序分析。