Title page for ETD etd-05152006-130213


Type of Document Master's Thesis
Author Buswell, Walter Scott
URN etd-05152006-130213
Title Expression of recombinant porcine preprorelaxin in Nicotiana tabacum
Degree Master of Science
Department Biological Systems Engineering
Advisory Committee
Advisor Name Title
Zhang, Chenming Mike Committee Chair
Agblevor, Foster Aryi Committee Member
Jelesko, John G. Committee Member
Keywords
  • recombinant protein
  • transgenic tobacco
  • relaxin
Date of Defense 2006-05-10
Availability unrestricted
Abstract
Relaxin is a small peptide hormone that has demonstrated potential therapeutic actions for cardiovascular disease and fibrosis. Additionally, relaxin has demonstrated the ability to protect the heart from injuries caused by ischemia and reperfusion, promote the healing of ischemic ulcers, and counteract allergic responses. The objective of this research was to express fully processed porcine relaxin in transgenic tobacco plants, as an alternative to current methods of producing relaxin.

Two recombinant relaxin genes were constructed that contained the patatin signal peptide cDNA fused in frame to prorelaxin cDNA, which was codon-optimized for expression in Nicotiana tabacum, under the control of either the “super” promoter or the dual enhanced cauliflower mosaic virus 35S promoter. Eighteen transgenic tobacco plants were generated that were transformed with the above recombinant genes. Preprorelaxin, mRNA was detected in 12 of the transgenic plants. Fully processed relaxin protein was not found in any tobacco plants that had demonstrated gene expression by northern blot analysis. Preprorelaxin was only identified in extracts from transgenic plants that contained the insoluble protein fraction, as determined by western blot analysis. Additionally, an increased yield of preprorelaxin was identified after incubation of tobacco leaves in an ubiquitin inhibitor.

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