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BIT 473/573: Protein Interactions

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:

  1. Introduction to protein interactions and realted thermodynamics
  2. In vivo vs. in vitro assays
  3. Use of EMSA, FP, ITC, SPR, AFM, and EM to study protein interactions
  4. Fluorescence microscopy and co-localization
  5. EcSSB protein properties and recent literature

Labs:

  1. Electrophoretic mobility shift assay (EMSA) of protein and DNA substrate to determine a dissociation constant
  2. BLI (BioLayer Interferometry) to verify the dissociation constant
  3. Chromatin immunoprecipitation (ChIP) to observe genomic localization of DNA-binding protein
  4. Fluorescence microscopy to see co-localization of labeled protein and DNA