Event Title

Expression and Purification of a Novel Calcium-Binding Protein, EfhX, Necessary for Phytopathogenesis in Xanthomonas Strains

Poster Number

045

Faculty Mentor

Jason Hurlbert, Ph.D.

College

College of Arts and Sciences

Department

Department of Chemistry, Physics, and Geology

Location

Richardson Ballroom – DiGiorgio Campus Center

Start Date

12-4-2019 2:15 PM

End Date

April 2019

Description

We have identified a gene in several strains of Xanthomonas that, when expressed, can elicit a hypersensitive response (hr) in tomato plants. Hr elicitation is normally limited to specific bacterial-host pairings, but introduction and expression of this gene by bacterial species that do not normally infect tomato serves to elicit hr, indicating that the encoded protein is crucial to the infectious process. Bioinformatic analysis of the protein, which we have named EfhX (EF-Hand containing protein from Xanthomonas) reveals that the protein is predicted to contain a single transmembrane alpha-helix, spanning amino acids 60 to 81, and two calcium binding domains, termed EF-Hands, in the carboxy-terminal domain of the protein. In order to better understand the function of this novel protein, we have cloned the EfhX gene from Xanthomonas aurantofolia and expressed it in Escherichia coli so that we can obtain quantities of the protein sufficient to grow protein crystals and determine the structure of the protein via X-ray diffraction. We have successfully purified the protein to homogeneity as determined by SDS-PAGE and anti-hexahistidine Western Blot.

Previously Presented/Performed?

SAEOPP McNair/SSS Scholars Research Conference, Atlanta, Georgia, June 2018

Grant Support?

Supported by an SC INBRE grant from the National Institute for General Medical Sciences (NIH-NIGMS), and by a Ronald E. McNair Post-Baccalaureate Achievement Program grant from the U.S. Department of Education

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Apr 12th, 2:15 PM Apr 12th, 4:15 PM

Expression and Purification of a Novel Calcium-Binding Protein, EfhX, Necessary for Phytopathogenesis in Xanthomonas Strains

Richardson Ballroom – DiGiorgio Campus Center

We have identified a gene in several strains of Xanthomonas that, when expressed, can elicit a hypersensitive response (hr) in tomato plants. Hr elicitation is normally limited to specific bacterial-host pairings, but introduction and expression of this gene by bacterial species that do not normally infect tomato serves to elicit hr, indicating that the encoded protein is crucial to the infectious process. Bioinformatic analysis of the protein, which we have named EfhX (EF-Hand containing protein from Xanthomonas) reveals that the protein is predicted to contain a single transmembrane alpha-helix, spanning amino acids 60 to 81, and two calcium binding domains, termed EF-Hands, in the carboxy-terminal domain of the protein. In order to better understand the function of this novel protein, we have cloned the EfhX gene from Xanthomonas aurantofolia and expressed it in Escherichia coli so that we can obtain quantities of the protein sufficient to grow protein crystals and determine the structure of the protein via X-ray diffraction. We have successfully purified the protein to homogeneity as determined by SDS-PAGE and anti-hexahistidine Western Blot.