Homeobox leucine zipper protein 9 (HAT9) -AT2G Homeodomain(s) -Leucine Zipper Motif -DNA Binding -Dimerization ?? ~ Helix-turn-helix 53 FWRV Exon 1 UTR Exon 3Exon 2 Intron 1Intron 3 Research Talk HC70AL Spring 2004 by Bobby Fam Evolutionary Relationships of HAT Proteins -Alignment of HAT9 and HAT22 reveals 90% identity in the homeodomain-leucine zipper region and 71% identity overall. -Little homology is seen outside of the homeodomain- leucine zipper region for other proteins. Gene Activity of AT2G kb kb Leaf: 1,2 Stem: 3,4 Flower: 5,6 Ovule: 7,8 Embryo: 9,10 Axis: 11,12 Leaf: 13,14 Flower: 15,16 -Gene specific bands for leaf, stem, flower, unfertilized ovules, and 30 DAP axis. -Arabidopsis: Activity in WT mature green seeds, WT Post- Maturation Green Seeds, and WT Roots -Possible Alternative Splicing 14 DAP embryo (lane 9). Madison Screen Round Control 1600 bp 1800 bp 4500 bp Round C 1kb -T-DNA insert in either superpool 10 or 22 -Sequencing data matches HAT9 but has no T-DNA -Reamplification yielded no matching bands SALK T-DNA Insertion Gene: AT5G67300SALK Line: WT H 2 0 1kb1kb WT H 2 0 FW/RV Gene-specific RV Gene-specifc & LBb1 primer -Plant 5 represents a homozygous T- DNA genotype -Plants 1,2,3,4,6,7,8, & 9 represent Phenotyping SALK Insert Plant Mutant FlowerWT leaf on left; Mutant leaf on right WT Silique top; Mutant Silique bottom Mutant Trichome -Mutant plant does not seem to differ in phenotype from the Wild-type in any significant manner To the Future -Sequence Madison Inserts -Identify DNA Pool (Reorder) -Identify Seed Pool -Identify Knock-out Line -Grow Plants & Observe AT2G22800 AT5G Investigate other knockout lines -Study other genes and their effects on seed development -Determine specific role of genes on seed development -Engineer the 21st Century Crop