Project Number: 5325-21000-008-00
Project Type: Appropriated

Start Date: Jan 20, 2002
End Date: Aug 31, 2006

Objective:
Identify endogenous cereal promoter elements that confer useful expression patterns within the plant, in a variety of tissues and with differing developmental specificities. Isolate and characterize regulatory sequences that confer expression in major organs but not in the seed grains used for food and feed. Develop molecular tools for integrating and deleting transgenic DNA within the genomes of cereal crops. Evaluate and adapt recombination systems for the precise placement of DNA into targeted locations, and for the subsequent selective removal of unneeded transgenic DNA from the genome. Test the hypothesis that the repeated stacking of DNA into a known site in the plant genome will result in predictable expression of each of the introduced transgenes.

Approach:
Use bioinformatics to scan EST and plant gene expression databases and hybridization of rice mRNA to cDNA arrays to identify candidate genes with novel expression patterns. Verify patterns by Northern hybridization. Isolate genomic DNA sequences for genes with useful expression patterns. Sequence flanking putative regulatory regions and fuse them to reporter gene coding sequences. Transform into cereal plants, initially rice. Characterize gene expression patterns in transgenic plants using reporter activity and in situ hybridization. Make results and sequences publicly available. Demonstrate activity of novel site-specific recombination systems in yeast and plant cells. Modify recombinases and/or recognition sequences to increase efficiencies in eukaryotic cells. Identify those systems that can mediate integration and deletions in plant cells. Test whether transgenes can be integrated into preexisting sites and whether they are fully expressed in those sites.

Project Team

Blechl, Ann Thilmony, Roger Thomson, James Ow, David

Project Annual Reports

FY 2003   FY 2002