Identification of Transcription Factor ZTF-4::GFP Binding Regions in L3

modENCODE_submission_3218 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: OP322(official name : ZTF-4 genotype : unc119(ed3);wgIs322(ztf-4::TY1 EGFP FLAG;unc119) outcross : 0 mutagen : None 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 spatio-temporal expression pattern of ZTF-4::EGFP fusion protein was examined through in vivo microscopy. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the ZTF-4 transcription factor. made_by : Bob Waterston''s lab from UW ); Developmental Stage: L3; Genotype: unc119(ed3);wgIs322(ztf-4::TY1 EGFP FLAG;unc119); Sex: Hermaphrodite; EXPERIMENTAL FACTORS: Developmental Stage L3; Target gene ztf-4; Strain OP322(official name : ZTF-4 genotype : unc119(ed3);wgIs322(ztf-4::TY1 EGFP FLAG;unc119) outcross : 0 mutagen : None 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 spatio-temporal expression pattern of ZTF-4::EGFP fusion protein was examined through in vivo microscopy. This strain was used for ChIP-seq experiments to map the in vivo binding sites for the ZTF-4 transcription factor. made_by : Bob Waterston''s lab from UW ); temp (temperature) 20 degree celsius

modENCODE_submission_3218 本提交源自迈克尔·斯奈德（Michael Snyder）主导的modENCODE项目。如需查阅modENCODE项目完整列表，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（C. elegans）中300种转录因子的DNA结合位点。将每一种转录因子基因与统一的绿色荧光蛋白（Green Fluorescent Protein, GFP）融合蛋白标签结合，可验证该基因在转基因动物中的时空表达模式是否正确。对每一株转基因线虫均采用抗GFP抗体进行染色质免疫沉淀（Chromatin Immunoprecipitation, ChIP）实验，结合的DNA通过Solexa GA2测序技术进行深度测序。关于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
生物样本来源：
菌株：OP322（官方命名：ZTF-4；基因型：unc119(ed3);wgIs322(ztf-4::TY1 EGFP FLAG;unc119)；回交次数：0；诱变剂：无；标签：GFP::3xFlag）
菌株构建说明：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米哈伊尔·萨罗夫（Mihail Sarov）利用托尼·海曼（Tony Hyman）的重组工程流程构建。所得质粒用于对unc-119(ed3)菌株进行基因枪法转化。通过活体显微镜成像检测了ZTF-4::EGFP融合蛋白的时空表达模式。该菌株被用于ChIP-seq实验，以绘制ZTF-4转录因子的体内结合位点。
构建方：来自华盛顿大学（UW）的鲍勃·沃特斯顿（Bob Waterston）实验室
发育阶段：L3期
基因型：unc119(ed3);wgIs322(ztf-4::TY1 EGFP FLAG;unc119)
性别：雌雄同体
实验因素：发育阶段L3；靶基因ztf-4；菌株OP322（官方命名：ZTF-4；基因型：unc119(ed3);wgIs322(ztf-4::TY1 EGFP FLAG;unc119)；回交次数：0；诱变剂：无；标签：GFP::3xFlag）
菌株构建说明：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米哈伊尔·萨罗夫（Mihail Sarov）利用托尼·海曼（Tony Hyman）的重组工程流程构建。所得质粒用于对unc-119(ed3)菌株进行基因枪法转化。通过活体显微镜成像检测了ZTF-4::EGFP融合蛋白的时空表达模式。该菌株被用于ChIP-seq实验，以绘制ZTF-4转录因子的体内结合位点。
构建方：来自华盛顿大学（UW）的鲍勃·沃特斯顿（Bob Waterston）实验室
培养温度：20摄氏度