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Homeobox leucine zipper protein 9 (HAT9) -AT2G22800
-Leucine Zipper Motif
-Homeodomain(s)
-Dimerization
-DNA Binding
-Helix-turn-helix
?? ~ 1400
FW
5’
RV
3’
1
102
UTR
5’
513
618
Exon 1
697
869
Exon 2
Intron 1
1203
Exon 3
Intron 2
Research Talk HC70AL Spring 2004 by Bobby Fam
1435
UTR
3’
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 homeodomainleucine 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 1kb
Leaf: 1,2
Stem: 3,4
Flower: 5,6
-Gene specific bands for leaf,
stem, flower, unfertilized
ovules, and 30 DAP axis.
Ovule: 7,8
Embryo: 9,10
Axis: 11,12
Leaf: 13,14
Flower: 15,16
-Possible Alternative Splicing 14 DAP embryo
(lane 9).
-Arabidopsis: Activity in
WT mature green
seeds, WT PostMaturation Green
Seeds, and WT Roots
Madison Screen
Round 1
Round 2
1
27
2 2
3
3 4
4
5
5 6
6 7 7
8 8
4500 bp
Control
10
1
1800 bp
22
1600 bp
-T-DNA insert in either
superpool 10 or 22
-Sequencing data matches
HAT9 but has no T-DNA
-Reamplification yielded no matching bands
9
10 C 1kb
SALK T-DNA Insertion
Gene: AT5G67300
FW/RV Gene-specific
1 2 3 4 5 6 7
8 9 WT H 20 1kb
SALK Line: 039074
RV Gene-specifc & LBb1 primer
1kb 1 2 3 4 5 6 7 8 9 WT H 20
-Plant 5 represents
a homozygous TDNA 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 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
-Sequence Madison Inserts
-Determine specific role of
genes on seed
development
-Identify DNA Pool (Reorder)
-Identify Seed Pool
-Identify Knock-out Line
-Grow Plants & Observe
-Engineer the 21st Century Crop