Molecular Biology

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Spring 2009 BI 425 UOG. Molecular Biology. Topics for Second Part of Molecular Biology Course: Structural Organization of Eukaryotic Chromosomes. NonTranscribed genes are less susceptible to Dnase 1 Digestion. Fluorescent Labeled Probes hybridized to Interphase Chromosomes.
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Spring 2009 BI 425 UOG Molecular Biology Topics for Second Part of Molecular Biology Course: Structural Organization of Eukaryotic Chromosomes NonTranscribed genes are less susceptible to Dnase 1 Digestion Fluorescent Labeled Probes hybridized to Interphase Chromosomes Prokaryotic Gene Regulation Chromosomes Deletion Analysis: Identify Upstream Transcription Control Sequences Determination of Consensus TATA Box Sequence Sequences Scanning Mutations Identify Transcription Control Elements Sequences General Pattern of Control Elements that Regulate Gene Expression in Yeast and other Eukaryotes. LIFE CYCLE OF HIV-1 Expression HIV Virus Latency DNA Provirus RNA Viral DNA Transcription Viral RNA RNA Processing Viral genomic RNA mRNA Virion Assembly Viral Proteins Maturation & Release of Virus if rev 3’ LTR vpu vpr pol env nef Shubir Ghosh, UCSF Shubir Ghosh, UCSF Expression Overview of Eukaryotic Transcriptional Control Expression Column Chromatography Separation of the three RNA Polymerases In Eukaryotes. Transcription Assay: Transcription Start Site Polymerases EMSA Polymerases Dnase 1 Footprinting Polymerases Dnase1 Footprint Assay in Transcription Factor Purification Polymerases Northern Blot & S1 Nuclease Assay: Polymerases Identify Enhancers of Transcription Northern Blot S1 Nuclease Assay Homework! In vitro Polymerases Transcription Assay: Identification of Transcription Factors In vivo Polymerases Transfection Assay: Identify & Evaluate Transcription Factors Eukaryotic Transcriptional Activators: Polymerases Features & Experimental Analysis Eukaryotic Transcriptional Activators: Polymerases Features & Experimental Analysis of GAL4 Eukaryotic Transcriptional Activators: Polymerases Features & Experimental Analysis of GAL4 Interaction of bacteriophage 434 Repressor with DNA Polymerases Interaction of Zinc Finger proteins with DNA Polymerases GR Interaction of Lecuine Zipper & basic HLH proteins with DNA Polymerases Yeast Gcn4 Transcription Activator Cooperative binding of two unrelated transcription factors to neighboring sites in a composite control element Combinatorial Possibilities due to formation of to Heterodimeric Transcription Factors Transcriptional Repression : Yeast mating Locus to Antibody and DNA probes colocalize RepressorProtein, Sir3, to with telomeric heterochromatin in Yeast nuclei Telomeres labeled by Hybridization to Telomere Specific DNA probe (Yellow) Telomere specific DNA Hybridization Probe Fluorescent labeled Antibody specific For Sir3 protein Schematic Model of Silencing Mechanism at Yeast Telomeres to Dependence of Silencing on Hypoacetylation of the Histone Tails Model of Transcriptional Activation Mechanism to See Handout on Histone Acetylation/Deacetylation! HACs & HDACs Chromatin Immunoprecipitation Assay: Evaluate State of Histone Acetylation on Nucleosome Transcriptional Activation by Gcn4 in Saccharomyces cerevisae Components of Gcn4 activated gene in yeast: Gcn4 Activated Gene in S.c UAS Gcn4 Gcn5 + Multisubunit Complex Mechanism of Transcriptional Activation by Gcn4 cerevisae Mechanism of Transcriptional Repression by Ume6 in Yeast cerevisae Components of Repression in yeast by Ume6: Ume6 Repressed gene in S.c Ume6 Multisubunit complex including: Sin3 Rpd3 Mechanism of Transcriptional Repression cerevisae Medistor Complex in Euksryotic Transcription cerevisae
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