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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:

  • 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