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Homeobox leucine zipper protein 9 (HAT9) -AT2G22800. -Leucine Zipper Motif. -Homeodomain(s). -Dimerization. -DNA Binding. -Helix-turn-helix. ?? ~ 1400. 1. 102. 513. 618. 697. 869. 1203. 1435. FW. RV. UTR. Exon 1. Exon 2. Exon 3. UTR. Intron 1. Intron 2. 5’. 3’. 5’. 3’.
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Homeobox leucine zipper protein 9 (HAT9) -AT2G22800 -Leucine Zipper Motif -Homeodomain(s) -Dimerization -DNA Binding -Helix-turn-helix ?? ~ 1400 1 102 513 618 697 869 1203 1435 FW RV UTR Exon 1 Exon 2 Exon 3 UTR Intron 1 Intron 2 5’ 3’ 5’ 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 AT2G22800 1kb 1 2 3 4 5 6 7 8910 11 12 13 14 15 16 1kb 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 1 Round 2 1 1 2 2 3 3 4 4 5 5 6 6 7 7 8 8 9 10 C 1kb 27 4500 bp Control 1800 bp 10 22 1600 bp -T-DNA insert in either superpool 10 or 22 -Reamplification yielded no matching bands -Sequencing data matches HAT9 but has no T-DNA
SALK T-DNA Insertion Gene: AT5G67300 SALK Line: 039074 FW/RV Gene-specific RV Gene-specifc & LBb1 primer -Plant 5 represents a homozygous T-DNA genotype 1 2 3 4 5 6 7 8 9 WT H20 1kb 1kb 1 2 3 4 5 6 7 8 9 WT H20 -Plants 1,2,3,4,6,7,8, & 9 represent
Phenotyping SALK Insert Plant WT Silique top; Mutant Silique bottom Mutant Flower WT leaf on left; Mutant leaf on right -Mutant plant does not seem to differ in phenotype from the Wild-type in any significant manner Mutant Trichome
To the Future… AT5G67300 -Investigate other knockout lines -Study other genes and their effects on seed development AT2G22800 -Determine specific role of genes on seed development -Sequence Madison Inserts -Identify DNA Pool (Reorder) -Engineer the 21st Century Crop -Identify Seed Pool -Identify Knock-out Line -Grow Plants & Observe