研究基因表达调控的基本方法 、 DNARNA与蛋白质的相互作用技术 Report gene Assay EMSA a ChIP SElEX Department of Biochemistry Molecular Biology e
Department of Biochemistry & Molecular Biology 6 研究基因表达调控的基本方法 一、DNA/RNA与蛋白质的相互作用技术 ❑ Report gene Assay ❑ EMSA ❑ ChIP ❑ SELEX
1. Reporter Gene Assay Measures the in vitro level of gene expression or transcriptional activity Transcription is initiated by binding of specific proteins to specific dna promoter regions Reporter assays use a specific DNA promoter fused to a reporter gene(luciferase, SEAP, GFP, etc.) Transcriptional activity of your promoter is measured y reporter expression Department of Biochemistry Molecular Biology e
Department of Biochemistry & Molecular Biology 7 Measures the in vitro level of gene expression or transcriptional activity Transcription is initiated by binding of specific proteins to specific DNA promoter regions Reporter assays use a specific DNA promoter fused to a reporter gene (luciferase, SEAP, GFP, etc.) Transcriptional activity of your promoter is measured by reporter expression 1. Reporter Gene Assay
Control of Gene Expression: Promoters Every gene has a promoter the dna sequence immediately surrounding the transcription start site. The promoter is the site where RNa polymerase and the so-called general transcription factors bind coactivators general transcription factors RNA polymerase TAtA box coding region promoter RNA transcript Department of Biochemistry Molecular Biology e
Department of Biochemistry & Molecular Biology 8 Control of Gene Expression: Promoters Every gene has a promoter, the DNA sequence immediately surrounding the transcription start site. The promoter is the site where RNA polymerase and the so-called general transcription factors bind
Report gene Assay RNA polymerase and Transcription factors Reporter gene Promoter Transcription mRNA Translation Reporter Protein Department of Biochemistry Molecular Biology e 9
Department of Biochemistry & Molecular Biology 9 Report gene Assay
R eport gene 报告基因是一种编码可被检测的蛋白质或酶的 基因,是一个其表达产物非常容易被检测或鉴 定的基因。通常将包含基因启动子的DNA片断 置于一个无关的缺乏内源性启动子活性的报告 基因编码序列的上游,通过检测报告基因的活 性或表达水平来间接度量启动子的活性。 Department of Biochemistry Molecular Biology e 10
Department of Biochemistry & Molecular Biology 10 Report gene 报告基因是一种编码可被检测的蛋白质或酶的 基因，是一个其表达产物非常容易被检测或鉴 定的基因。通常将包含基因启动子的DNA片断 置于一个无关的缺乏内源性启动子活性的报告 基因编码序列的上游，通过检测报告基因的活 性或表达水平来间接度量启动子的活性