|
OBJECTIVES
Our overall goal is to use genomic approaches to
drive an increased understanding of the network of enzymatic and other
processes involved in the production of seed TAGs, and in particular
how these processes have coevolved to facilitate the synthesis and
accumulation of unusual fatty acids.
Our specific objectives under
this grant are to:
- Use
biochemical, molecular-genetic and bioinformatic techniques to
investigate the primary enzymes of fatty acid modification for
each of our model systems. Although some
of these enzymes have already been studied by us and others, it is
clear that we need to do more to understand the potential deficiencies
of these enzymes in a transgenic context particularly as they relate
to protein stability and interaction with other pathway components
in the ER membrane.
- Build
and extend genomic resources, through 454 sequencing and other
techniques, to provide comprehensive access to all the gene candidates
that may impact the efficiency with which hydroxy, conjugated,
and cyclopropane fatty acids are mobilized through the possible
reaction paths into TAG.
- Systematically
evaluate these candidate genes and proteins using coexpression studies, in
vivo labeling experiments, enzyme assays and reverse-genetic
approaches that will specify their individual contributions.
- Test
for synergistic effects provided by combining genes in poly-transgenic
lines. The information
obtained will then be integrated with results from proteomics analysis
and experimental tests of protein-protein interaction using split-ubiquitin
assays to model the biosynthetic pathways within the broader cellular
context.
- Broaden
public understanding and excite young students about the potential
of plants to serve as the chemical factories of the future. Train
current and future plant scientists to apply genomics in a focused
way to solve long-standing problems in understanding complex biosynthetic
pathways in plants and to appreciate how these pathways may have
evolved during angiosperm radiation.
|
