Homeobox leucine zipper protein 9 (HAT9) -AT2G22800
-Homeodomain(s) -Leucine Zipper Motif
-DNA Binding -Dimerization
?? ~ 1400
-Helix-turn-helix
5’ 3’
FW RVExon 1UTR UTRExon 3Exon 2
Intron 1 Intron 2
102 513 618 697 869 1203 14351
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 8 9 10 11 12 13 14 15 16 1kbLeaf: 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
10 22
27
Control
1600 bp
1800 bp
4500 bp
Round 2
1 1 2 2 3 3 4 4 5 5 6 6 7 7 8 8 9 10 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 InsertionGene: AT5G67300 SALK Line: 039074
1 2 3 4 5 6 7 8 9 WT H20 1kb 1kb 1 2 3 4 5 6 7 8 9 WT H20
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 Flower WT 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
AT5G67300
-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