Identification of Transcription Factor UNC-62::GFP Binding Regions in L1. Caenorhabditis elegans

modENCODE_submission_3375 This submission comes from a modENCODE project of Michael Snyder. For full list of modENCODE projects, see http://www.genome.gov/26524648 Project Goal: We are identifying the DNA binding sites for 300 transcription factors in C. elegans. Each transcription factor gene is tagged with the same GFP fusion protein, permitting validation of the gene's correct spatio-temporal expression pattern in transgenic animals. Chromatin immunoprecipitation on each strain is peformed using an anti-GFP antibody, and any bound DNA is deep-sequenced using Solexa GA2 technology. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Overall design: EXPERIMENT TYPE: CHIP-seq. BIOLOGICAL SOURCE: Strain: OP600(official name : OP600 genotype : unc119(ed3);wgIs600(unc-62::TY1 EGFP FLAG;unc119) outcross : 3 mutagen : Bombard tags : GFP::3xFlag description : This strain's transgene was constructed by Mihail Sarov at the Max Planck Institute for Cell Biology in Tubiginen using Tony Hyman's recombineering pipeline. The resulting plasmid was used for biolistic transformation of an unc-119(ed3) strain. The UNC-62::EGFP fusion protein is expressed in the correct unc-62 spatio-temporal expression pattern. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the UNC-62 transcription factor. made_by : S Kim ); Developmental Stage: fed L1; Genotype: unc119(ed3);wgIs600(unc-62::TY1 EGFP FLAG;unc119); Sex: Hermaphrodite; EXPERIMENTAL FACTORS: Developmental Stage fed L1; Target gene unc-62; Strain OP600(official name : OP600 genotype : unc119(ed3);wgIs600(unc-62::TY1 EGFP FLAG;unc119) outcross : 3 mutagen : Bombard tags : GFP::3xFlag description : This strain's transgene was constructed by Mihail Sarov at the Max Planck Institute for Cell Biology in Tubiginen using Tony Hyman's recombineering pipeline. The resulting plasmid was used for biolistic transformation of an unc-119(ed3) strain. The UNC-62::EGFP fusion protein is expressed in the correct unc-62 spatio-temporal expression pattern. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the UNC-62 transcription factor. made_by : S Kim ); temp (temperature) 20 degree celsius

modENCODE_submission_3375 本数据集提交源自Michael Snyder主持的modENCODE项目。完整modENCODE项目列表请参阅：http://www.genome.gov/26524648。
项目目标：本项目旨在鉴定秀丽隐杆线虫（C. elegans）中300种转录因子的DNA结合位点。每个转录因子基因均携带相同的绿色荧光蛋白（GFP）融合标签，可用于验证转基因动物中该基因的正确时空表达模式。我们通过抗GFP抗体对每株菌株开展染色质免疫沉淀实验，结合的DNA采用Solexa GA2技术进行深度测序。数据使用条款与规范请参考：http://www.genome.gov/27528022 及 http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf。
总体实验设计：
实验类型：染色质免疫沉淀测序（ChIP-seq）。
生物样本来源：菌株OP600（官方名称：OP600；基因型：unc119(ed3);wgIs600(unc-62::TY1 EGFP FLAG;unc119)；回交次数：3次；诱变方式：基因轰击；标记：GFP::3xFlag）。该菌株的转基因载体由德国蒂宾根马克斯·普朗克细胞生物学研究所的Mihail Sarov利用Tony Hyman的重组工程平台构建，所得质粒用于unc-119(ed3)菌株的基因轰击转化。UNC-62::EGFP融合蛋白的表达模式与unc-62基因的天然时空表达模式一致，由S Kim构建并用于ChIP-seq实验，以绘制UNC-62转录因子的体内结合位点。
发育阶段：喂食期L1幼虫；
基因型：unc119(ed3);wgIs600(unc-62::TY1 EGFP FLAG;unc119)；
性别：雌雄同体。
实验影响因素：发育阶段为喂食期L1幼虫；靶基因为unc-62；实验菌株为OP600（官方名称：OP600；基因型：unc119(ed3);wgIs600(unc-62::TY1 EGFP FLAG;unc119)；回交次数：3次；诱变方式：基因轰击；标记：GFP::3xFlag）。
培养温度：20摄氏度。