East Tennessee State University
cDNA Cloning, Expression and Characterization of a Putative Glucosyltransferase (GT) from Grapefruit (<em>Citrus paradisi</em>) Leaves.
Abstract
dc:description.abstract<p>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-<sup>14</sup>C-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.</p>
Degree
thesis:*- Name thesis:degree_name
- MS (Master of Science)
- Level thesis:degree_level
- Thesis - restricted
- Discipline thesis:degree_discipline
- Biology
- Year dc:date.issued
- 2004
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Roy Sarkar, Tapasree
Subjects
dc:subject × 7Rights
dc:rights- Statement dc:rights
-
- Copyright by the authors.
Identifiers
dc:identifier.*- Repository record dc:identifier
- https://dc.etsu.edu/etd/908
- OAI identifier oai:identifier
- oai:dc.etsu.edu:etd-2065