Discipline: Biological Sciences
Subcategory: Plant Research
James Taylor - Fort Valley State University
Co-Author(s): Abhaya Dandekar, Ph.D , University of California, Davis, CA
Anthrax toxin receptor- mediated drug development for blocking anthrax toxin has increased in popularity the past ten years. During this study, we worked on producing a secreted anthrax decoy fusion protein comprised of capillary morphogenesis gene-2 (CMG2). CMG2 functions to capture bacteria during photogenesis, and can play a major role in anthrax photogenesis. Using the cauliflower Mosaic Virus 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to get high amounts of recombinant CMG2-Fc-Apo protein accumulation in walnut embryos. We observed the embryos over eight days post infiltration. The day of maximum production was day 6. Affinity chromatography purification of CMG2 protein from the embryo and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis. Mass spectrometry analysis confirmed molecular integrity of the secreted protein. Altogether these findings demonstrate that high level production of CMG2 can be achieved by transient production in walnut embryos with apoplast targeting.Not Submitted
Funder Acknowledgement(s): This work was supported by NSF HRD (#1238789)"HBCU-UP" Targeted Infusion grant to Prof. Seema Dhir at Fort Valley State University, Fort Valley, GA.
Faculty Advisor: Prof. Seema Dhir, firstname.lastname@example.org
Role: I first expressed the recombinant CMG2-Fc-Apo protein in Walnut embryos. Then I performed affinity chromatography of the protein to purify it as well as Mass Spectrophotometry to further confirm the molecular integrity of the protein.