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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/24278

Title: Optimization of Purification Conditions for the Pseudomonas syringae HopZ1a Type III Secreted Effector Protein for Structural and Functional Studies
Authors: Quach, Van Chau
Advisor: Christendat, Dinesh
Desveaux, Darrell
Department: Cell and Systems Biology
Keywords: Molecular biology
microbiology
Issue Date: 6-Apr-2010
Abstract: HopZ1a is a type III secreted effector (TTSE) protein from Pseudomonas syringae. The goal of this study was to obtain a 3D crystal structure of HopZ1a to provide insight into its biochemical function. The first objective was to obtain HopZ1a protein that was sufficiently abundant and pure for crystallographic studies. Purification conditions were optimized and multiple constructs of HopZ1a were generated using secondary structure prediction programs as well as structural characteristics inherent to TTSEs. Truncations of HopZ1a from the N- and C-terminus led to a soluble, proteolytically resistant construct, HopZ1a66-261. This protein formed granular precipitates in crystallography screens. These conditions will provide the basis for refinement screens aimed at optimizing the HopZ1a crystallization conditions. Overall, the soluble constructs described in this study will provide invaluable tools for future in vitro functional and structural studies of this important family of type III secreted effector proteins.
URI: http://hdl.handle.net/1807/24278
Appears in Collections:Master
Department of Cell and Systems Biology - Master theses

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