Author (Your Name)

Natalie Wayne, Colby College

Date of Award

2006

Document Type

Honors Thesis (Open Access)

Department

Colby College. Chemistry Dept.

Advisor(s)

Russell R. Johnson

Second Advisor

Frederick LaRiviere

Abstract

Abscisic acid (ABA)-mediated gene expression is a critical component of plant responses to this important hormone, which affects plant growth, development, and responses to environmental stresses. Plant responses to ABA are mediated by a number of factors including PKABA 1> an ABA-induced protein kinase involved in ABA-suppressed gene expression in cereal grains, and TaWD40, which has previously been shown to physically interact with PKABA1. A full-length 1.9 kb TaWD40 cDNA, CK210682, was sequenced as part of this project. Based on the deduced protein sequence, it is thought that TaWD40 may belong to the family of E3 ubiquitin ligases, possibly targeting PKABA 1 for destruction. Construction of expression plasmids for overproduction of the TaWD40 polypeptide in E. coli is currently underway. The TaWD40 cDNA has been successfully amplified from the source plasmid and inserted into an intermediate plasmid, peR2.I. The TaWD40 cDNA is currently being cloned from the peR2.l intermediate plasmid into two different expression vectors, pRSET-A and pMAL-c2x, for future protein production and purification.

Keywords

Wheat grain, PKABA1, TaWDR40, E3 ubiquitin ligases, protein production, purification

Included in

Chemistry Commons

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