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Full Description
This volume provides a comprehensive set of protocols that can be used by any research lab to investigate diverse functional and structural properties of Single Stranded DNA Binding Proteins (SSBs) from eubacterial, archaeal, eukaryotic, mitochondrial and viral systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Single Stranded DNA Binding Proteins aims to be a useful practical guide to researchers to help further their study in this field.
Contents
The Essential, Ubiquitous Single-Stranded DNA-binding Proteins.- Single-stranded DNA Binding Proteins in the Archaea.- Single-molecule Fluorescence Methods to Study Protein Exchange Kinetics in Supramolecular Complexes.- Comparing SSB-PriA Functional and Physical Interactions in Gram-positive and -Negative Bacteria.- In vivo binding of Single-stranded DNA-binding Protein to Stalled Replication Fork Helicases.- Magnetic Tweezers-based Single Molecule Assays to Study Interaction of E. coli SSB with DNA and RecQ Helicase.- High-throughput Screening to Identify Inhibitors of SSB-Protein Interactions.- Single-Molecule Tethered Particle Motion Studies on the DNA Recombinase Filament Assembly and Disassembly.- Generation of Fluorescent Versions of Saccharomyces cerevisiae RPA to Study the Conformational Dynamics of its ssDNA-binding Domains.- RPA-1 from Leishmania sp.: Recombinant Protein Expression and Purification, Molecular Modeling and Molecular Dynamics Simulations Protocols.- Single-stranded DNA Curtains for Single-molecule Visualization of Rad51-ssDNA Filament Dynamics.- Following Trypanosoma cruzi RPA-DNA Interaction Using Fluorescent in Situ Hybridization Coupled with Immunofluorescence (FISH/IF).- Quantifying the affinity of Trypanosoma cruzi RPA-1 to the Single-stranded DNA Overhang of the Telomere using Surface Plasmon Resonance.- Expression, Purification, and Solution-State NMR Analysis of the Two Human Single-stranded DNA Binding Proteins hSSB1 (NABP2/OBFC2B) and hSSB2 (NAPB1/OBFC2A).- Atomic Force Microscopy Reveals that the Drosophila Telomere-Capping Protein Verrocchio is a Single-stranded DNA Binding Protein.- Analysis of Mitochondrial SSB-DNA Complexes and their Effects on DNA Polymerase γ Activity by Electron Microscopy and Enzymatic Assays.- Optical Tweezers to Investigate the Structure and Energetics of Single-Stranded DNA-binding Protein-DNA Complexes.- Measurements of Real-time Replication Kinetics of DNA Polymerases on ssDNA Templates Coated with Single-stranded DNA-binding Proteins.- Selective Suppression of Endogenous Gene Expression Using RNAi in Drosophila Schneider S2 cells.- Stimulation of Variant forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-stranded DNA-binding Protein.- Measuring the Complex Effects of the Single-Stranded DNA-Binding Protein gp2.5 on Primer Synthesis and Extension by the Bacteriophage T7 Replisome.- Strand Displacement and Unwinding Assays to Study the Concerted Action of the DNA Polymerase and SSB during Phi29 TP-DNA rReplication.- Structural Characterization of a Single-stranded DNA-binding Protein: A Case Study of the ORF6 protein from Bacteriophage Enc34.
