Determining The 3-Dimensional Folding Structure Of A Citrus Cankar Causing Protein Produced By The Xanthomonas Axonopodis Bacteria

School Name

Governor's School for Science and Math

Grade Level

12th Grade

Presentation Topic

Biochemistry

Presentation Type

Mentored

Mentor

Mentor: Dr. Hurlbert; Department of Chemistry, Biochemistry, Physics, and Geology, Winthrop University

Oral Presentation Award

3rd Place

Abstract

A protein produced by the Xanthomas axanopodis bacterium causes citrus cankar. The determination of the 3-dimensional folding structure of this protein would be a key step in developing a cure for this disease. To determine the 3-dimensional folding structure of a protein, the protein must first be produced in sufficient quantities. The goal of this research was to identify an appropriate strain of E. coli to produce the protein and to develop a purification protocol so that, in later experiments, the structure could be found by performing X-ray crystallography. Several transformed strains of E. coli were tested for their ability to manufacture the protein and E. coli Rosetta 2 was found to produce the highest levels. A Fast Protein Liquid Chromatography (FPLC) purification protocol was developed and a 38 kDa band of the putative protein was identified. This band has the same molecular weight as the Xanthomas Axanopodis protein. Future work would include verifying the identity of this protein band by amino acid sequencing and the optimization of this protocol to produce sufficient quantities for crystallography.

Location

Owens 203

Start Date

4-16-2016 11:15 AM

COinS
 
Apr 16th, 11:15 AM

Determining The 3-Dimensional Folding Structure Of A Citrus Cankar Causing Protein Produced By The Xanthomonas Axonopodis Bacteria

Owens 203

A protein produced by the Xanthomas axanopodis bacterium causes citrus cankar. The determination of the 3-dimensional folding structure of this protein would be a key step in developing a cure for this disease. To determine the 3-dimensional folding structure of a protein, the protein must first be produced in sufficient quantities. The goal of this research was to identify an appropriate strain of E. coli to produce the protein and to develop a purification protocol so that, in later experiments, the structure could be found by performing X-ray crystallography. Several transformed strains of E. coli were tested for their ability to manufacture the protein and E. coli Rosetta 2 was found to produce the highest levels. A Fast Protein Liquid Chromatography (FPLC) purification protocol was developed and a 38 kDa band of the putative protein was identified. This band has the same molecular weight as the Xanthomas Axanopodis protein. Future work would include verifying the identity of this protein band by amino acid sequencing and the optimization of this protocol to produce sufficient quantities for crystallography.