Identification of Transcription Factor ALY-2::GFP Binding Regions in L3. Caenorhabditis elegans

modENCODE_submission_3382 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: OP217(official name : OP217 genotype : unc119(ed3);wgIs217(aly-2:TY1 EGFP FLAG;unc119) outcross : 3 transgene : aly-2 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 ALY-2::EGFP fusion protein is expressed in the correct aly-2 spatio-temporal expression pattern. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the ALY-2 transcription factor. made_by : Mihail Sarov ); Developmental Stage: L3; Genotype: unc119(ed3);wgIs217(aly-2:TY1 EGFP FLAG;unc119); Sex: Hermaphrodite; Transgene: aly-2; EXPERIMENTAL FACTORS: Developmental Stage L3; Target gene aly-2; Strain OP217(official name : OP217 genotype : unc119(ed3);wgIs217(aly-2:TY1 EGFP FLAG;unc119) outcross : 3 transgene : aly-2 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 ALY-2::EGFP fusion protein is expressed in the correct aly-2 spatio-temporal expression pattern. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the ALY-2 transcription factor. made_by : Mihail Sarov ); temp (temperature) 20 degree celsius

modENCODE_submission_3382 本提交来自Michael Snyder主导的模式生物DNA元件百科全书（modENCODE）项目。如需查看完整modENCODE项目列表，请访问http://www.genome.gov/26524648。
项目目标：本研究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。将每种转录因子基因与统一的绿色荧光蛋白（Green Fluorescent Protein, GFP）融合蛋白标签融合，以此验证转基因动物中目标基因的正确时空表达模式。对每株转基因线虫采用抗GFP抗体进行染色质免疫共沉淀（Chromatin Immunoprecipitation, ChIP），所富集的结合DNA通过索莱克斯GA2（Solexa GA2）测序技术进行深度测序。
关于数据使用的条款与条件，请参阅http://www.genome.gov/27528022及http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf。
整体实验设计：
实验类型：ChIP-seq（染色质免疫共沉淀测序）
生物样本来源：
菌株：OP217（官方名称：OP217；基因型：unc119(ed3);wgIs217(aly-2:TY1 EGFP FLAG;unc119)；回交次数：3；转基因片段：aly-2；标签类型：Bombard；融合标签：GFP::3xFlag）
菌株描述：该菌株的转基因片段由蒂宾根马克斯·普朗克细胞生物学研究所的Mihail Sarov借助Tony Hyman的重组工程流程构建所得，将获得的质粒用于unc-119(ed3)菌株的基因枪转化。ALY-2::EGFP融合蛋白的表达模式与aly-2基因的天然时空表达模式一致。该菌株被用于ChIP-seq实验以定位ALY-2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);wgIs217(aly-2:TY1 EGFP FLAG;unc119)；性别：雌雄同体；转基因片段：aly-2
实验影响因素：发育阶段为L3期；靶基因为aly-2；所用菌株为OP217（官方名称：OP217；基因型：unc119(ed3);wgIs217(aly-2:TY1 EGFP FLAG;unc119)；回交次数：3；转基因片段：aly-2；标签类型：Bombard；融合标签：GFP::3xFlag）
菌株描述：该菌株的转基因片段由蒂宾根马克斯·普朗克细胞生物学研究所的Mihail Sarov借助Tony Hyman的重组工程流程构建所得，将获得的质粒用于unc-119(ed3)菌株的基因枪转化。ALY-2::EGFP融合蛋白的表达模式与aly-2基因的天然时空表达模式一致。该菌株被用于ChIP-seq实验以定位ALY-2转录因子的体内结合位点。构建者：Mihail Sarov
培养温度：20摄氏度