User:Jrfisher

Bio
2002 B.S. Biochemistry, University of New Hampshire (UNH)

"Two-Hybrid Analysis of LRR-Binding Peptides"

2004 M.S. Biochemistry and Molecular Biology, UNH,

"Isolation and Characterization of Leucine-Rich Repeat (LRR) Binding Peptides"

Intro
Technical assistant and safety representative, for the Keating lab since summer of 2004. I mainly troubleshoot and optimize molecular biology, protein purification and protein binding experiments.

Research
I.) Explore High-Throughput techniques of protein purification

A vacuum manifold coupled with a series of 96 well filter plates is used to control the flow of dual affinity tagged proteins over corresponding affinity resins

Proteins are then transferred to a UV acceptable 96 well plate and lyophilized. This method permits the use of any buffer to re-suspend proteins and determine concentration

The 96 well of pure proteins can then be used to perform experiments to analyze protein-protein interactions

PHOTO of APPARATUS

Millipore Multi-screen HTS vacuum manifold

II.) Bcl-2/BH3 Interaction Profiles and Specificity Determinants

Purify a panel of human and murine BH3-only peptides and mutants to test binding specificities against a subset of Bcl-2 family proteins. Identify amino acids important for various interactions.

III.) Identify and Characterize viral bZIP and human bZIP coiled coil interactions Purify several human bZIP’s and viral bZIP’s coiled coil regions to experimentally determine and characterize homo and hetero-dimeric interactions. Generate a series of mutations and designed coiled coil peptides to identify key binding residues and the biochemical composition of interaction interfaces.