Identification of Transcription Factor DAF-12::GFP Binding Regions in L3. Caenorhabditis elegans

modENCODE_submission_3381 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: OP222(official name : OP222 genotype : unc119(ed3);wgIs222(daf-12:TY1 EGFP FLAG;unc119) outcross : 3 transgene : daf-12 tags : Bombard tag : 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 DAF-12::EGFP fusion protein is expressed in the correct daf-12 spatio-temporal expression pattern. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the DAF-12 transcription factor. made_by : Mihail Sarov ); Developmental Stage: L3; Genotype: unc119(ed3);wgIs222(daf-12:TY1 EGFP FLAG;unc119); Sex: Hermaphrodite; Transgene: daf-12; EXPERIMENTAL FACTORS: Developmental Stage L3; Target gene daf-12; Strain OP222(official name : OP222 genotype : unc119(ed3);wgIs222(daf-12:TY1 EGFP FLAG;unc119) outcross : 3 transgene : daf-12 tags : Bombard tag : 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 DAF-12::EGFP fusion protein is expressed in the correct daf-12 spatio-temporal expression pattern. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the DAF-12 transcription factor. made_by : Mihail Sarov ); temp (temperature) 20 degree celsius

modENCODE_submission_3381 本提交源自Michael Snyder主持的modENCODE（Model Organism Encyclopedia of DNA Elements）项目。如需获取完整modENCODE项目列表，请访问http://www.genome.gov/26524648。
项目目标：本项目旨在鉴定秀丽隐杆线虫（Caenorhabditis elegans，C. elegans）中300种转录因子的DNA结合位点。每一种转录因子基因均融合了相同的绿色荧光蛋白（Green Fluorescent Protein, GFP）标签，以此验证该基因在转基因动物中的时空表达模式是否正确。研究人员针对每一株转基因菌株，采用抗GFP抗体开展染色质免疫共沉淀实验，随后通过Solexa GA2测序技术对结合的DNA片段进行深度测序。有关数据使用的条款与规范，请参阅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：ChIP-seq（染色质免疫共沉淀测序）
生物样本来源：
菌株：OP222（官方名称：OP222；基因型：unc119(ed3);wgIs222(daf-12:TY1 EGFP FLAG;unc119)；回交次数：3次；转基因片段：daf-12；标签：Bombard标签：GFP::3xFlag）
样本描述：该菌株的转基因片段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的重组工程流程构建所得。所得质粒被用于对unc-119(ed3)菌株进行生物弹道转化。DAF-12::EGFP融合蛋白的时空表达模式与内源daf-12基因完全一致。该菌株被用于ChIP-seq实验，以定位DAF-12转录因子的体内结合位点。构建者：Mihail Sarov
发育阶段：L3期；基因型：unc119(ed3);wgIs222(daf-12:TY1 EGFP FLAG;unc119)；性别：雌雄同体；转基因片段：daf-12
实验影响因素：发育阶段L3期；靶基因daf-12；菌株OP222（官方名称：OP222；基因型：unc119(ed3);wgIs222(daf-12:TY1 EGFP FLAG;unc119)；回交次数：3次；转基因片段：daf-12；标签：Bombard标签：GFP::3xFlag）
样本描述：该菌株的转基因片段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的重组工程流程构建所得。所得质粒被用于对unc-119(ed3)菌株进行生物弹道转化。DAF-12::EGFP融合蛋白的时空表达模式与内源daf-12基因完全一致。该菌株被用于ChIP-seq实验，以定位DAF-12转录因子的体内结合位点。构建者：Mihail Sarov
培养温度：20摄氏度