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Overview:
- Participants will be introduced to a variety of methods for studying the interactions of proteins, with other proteins and various substrates, and the functions of these interactions inside cells.
- Students will get hands-on experience with a variety of complimentary in vitro and in vivo techniques to study the interactions of proteins with DNA and other proteins.
Lecture topics:
- Introduction to protein interactions and realted thermodynamics
- In vivo vs. in vitro assays
- Use of EMSA, FP, ITC, SPR, AFM, and EM to study protein interactions
- Fluorescence microscopy and co-localization
- EcSSB protein properties and recent literature
Labs:
- Electrophoretic mobility shift assay (EMSA) of protein and DNA substrate to determine a dissociation constant
- BLI (BioLayer Interferometry) to verify the dissociation constant
- Chromatin immunoprecipitation (ChIP) to observe genomic localization of DNA-binding protein
- Fluorescence microscopy to see co-localization of labeled protein and DNA
