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Degree Name
MS (Master of Science)
Program
Biology
Date of Award
8-2004
Committee Chair or Co-Chairs
Cecilia A. McIntosh, Lee M. Pike
Committee Members
Hugh A. Miller III
Abstract
Flavonoids are plant secondary metabolites that are integral to our lives. Grapefruits are well-known for production of unique glucosylated products and the enzymes responsible are UDP-glucose:glucosyltransferases (GTs). The objective of this research was to obtain full-length clones of putative grapefruit GTs, express them, and characterize them. Previously, gene specific primers (from conserved PSPG box) and clone specific primers (from partial 5' clones) were designed, and a compiled sequence attained using SMART RACE RT-PCR. A full-length clone was obtained using primers designed from the extreme ends of the compiled sequence. The full-length clone was inserted into expression vector (pET32a) and transformed into expression host BL21(DE3)RIL. Expressed protein was tested for GT activity using different flavonoid aglycones and UDP-14C-glucose as glucose donor. Results indicated that the expressed protein was probably not a flavonoid GT. A directionally cloned grapefruit leaf cDNA library is undergoing EST mining to identify additional GT candidates.
Document Type
Thesis - restricted
Recommended Citation
Roy Sarkar, Tapasree, "cDNA Cloning, Expression and Characterization of a Putative Glucosyltransferase (GT) from Grapefruit (Citrus paradisi) Leaves." (2004). Electronic Theses and Dissertations. Paper 908. https://dc.etsu.edu/etd/908
Copyright
Copyright by the authors.