Title page for ETD etd-04272001-004931


Type of Document Dissertation
Author Santos, Michael Carmelo Orda
Author's Email Address mike.santos@vt.edu
URN etd-04272001-004931
Title Immunomodulation of Flavonoid Biosynthesis in Transgenic Arabidopsis thaliana
Degree PhD
Department Biology
Advisory Committee
Advisor Name Title
Winkel, Brenda S. J. Committee Chair
Beers, Eric P. Committee Member
Rutherford, Charles L. Committee Member
Stevens, Ann M. Committee Member
Walker, Richard A. Committee Member
Keywords
  • plant metabolic engineering
  • scFv
  • flavonoids
Date of Defense 2001-04-10
Availability unrestricted
Abstract
In an effort to test the feasibility of intracellular expression of enzyme-targeted antibodies to alter metabolism, recombinant antibody fragments in the single-chain format (scFv) were isolated from a phage display library using Arabidopsis chalcone isomerase (CHI) of the flavonoid biosynthetic pathway as the antigen. Each of the genes encoding the scFv’s was cloned into a plant transformation vector, which was subsequently used to generate transgenic plants. One transgenic line with low expression of one of the scFv’s appeared to have an altered flavonoid metabolism, as evidenced by a reduced capacity for anthocyanin accumulation and a reduction in flavonol glycosides in seedlings. Strong corroborating evidence that implicated the binding of scFv to CHI in the phenotypic alterations was obtained from protein mobility shift assays. Taken together, the results indicate that scFv-mediated metabolic alteration is possible in plants. Thus, we show that intracellular expression of scFv’s can be exploited as an additional tool for metabolic engineering.
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