Date of Award

5-2018

Document Type

Thesis

Department

Environmental and Forest Biology

Thesis Advisor

Lee Newman

Abstract

The use of injectable vaccines and pharmaceuticals pose logistical problems in many parts of the world. This problem has previously been investigated using various plants to produce pharmaceuticals, which would eliminate the problem of injection sterility. However, concentration determination in the harvested plants still pose a critical problem. For other medicines that might be cytotoxic, there is the additional problem that production in the living plant tissues could kill the plant before they could produce enough compound to be economically viable. The solution proposed by this project focused on the use of the promotor for a 58 kD protein in Capsicum to target pharmaceutical production to the ripened fruit of the red pepper plant. Due to time restraints in the lab this experiment and paper will focus on the isolation which can then be used in the future to begin work on the promotors. This avoids the issue of cytotoxicity as the promotor drives protein synthesis only in the final ripening states of the fruit, which can be harvested when ripened, thus preventing the entire plant from dying. These peppers can be harvested, ground and dried, providing a sample that can be tested for concentration and uniformly allocated into capsules. The protein has thus far been isolated from the chromoplast of the Capsicum fruit using a sucrose gradient centrifugation using a modified method for plant isolations. The next step will be the isolation of both mRNA and genomic DNA to search for the promoter regions using a cDNA clone for the protein of interest. Once the promoter regions are identified a DNA cassette can be produced containing a marker gene and inserted into a range of fruit bearing plants to determine which fruits produce the highest level of the marker, and also the timing of the marker. The final goal would be the insertion of a pharmaceutically relevant gene into the DNA cassette, and create transgenic plants that produce the desired compound only at the time of ripening thus preventing the entire plant being killed.

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