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