Arabidopsis Thaliana Gene AT5G58610

1
Arabidopsis Thaliana GeneAT5G58610

• Nancy Gallardo
• HC70AL

2
What Protein Does AT5G58610 Code for?
PHD-Type Zinc Finger Transcription Factor

• Regulates gene expression
• 1065 amino acids

3
What's Around Gene AT5G58610 ?
AT5G58600
5'
3'
3' 5' 3'
5' 5' 3'
AT5G8610
AT5G58620
AT5G58600
AT5G58620
5' 3' 5'
3' 3' 5'

5'
3'
1045bp 2286bp
23703997-23708279
4
What is the Structure of AT5G58610 ?
3' 5'
5' 3'
Exon
Exon
Exon
Exon
Exon
Exon
Exon
Exon
Exon

• Chromosome 5
• 4283 base pairs
• Reverse Orientation
• 9 Exons and 8 Introns

5
Expected T-DNA InsertionSalk_041367
LBb1-150bp
Expected Sizes WT 908bp T-DNA 696bp
T-DNA
Rv- 20bp
3'
5'
Intron 2
Intron 1
546bp 362bp
Exon 1
Fw- 20bp
6
What Were the Genotyping Results ?
WT 908 T-DNA696
PCR Genotyping AT5G58610
700bp
1 agarose, 105 Volts, 04/15/2008, 1.5Hour Run
7
Do these results agree with the RT-PCR results?
8
Where is AT5G58610 Active?
RT-PCR AT5G58610
1 Agarose 1 Hour Run 70 Volts 04/24/2008
9
Arabidopsis Thaliana GeneAT4G33280
10
What Protein Does My Gene Code for?

• B3-Domain Transcription Factor Families
• Regulate gene expression
• 3 families in Arabidopsis Thaliana
• 337 amino acids

11
What's Around Gene AT4G33280 ?
5'
3'
3' 3'
5'
AT4G33280
AT4G33290
AT4G33270
5'
5' 3'
561bp
217bp
3'
5'
16047358-16049359
12
What is the Structure of AT4G33280?
3' 5'
5' 3'
Exon
Exon
Exon
Exon
Exon
UTR
UTR
Intron Intron
Intron Intron

• Chromosome 4
• 2002 Base pairs
• Reverse Orientation
• 5 Exons, 4 Introns, 2 UTRs

13
Expected T-DNA Insertion
Salk_072801
LBb1-150bp
Expected Sizes WT 402bp T-DNA 292bp
T-DNA
Rv-20bp


5'
3'
AT4G32270
Intron 4
260Bp 142bp
Exon 5
Fw-23bp
14
What Were the Plants' Genotypes ?
Expected T-DNA292bp Expected Wild
Type402bp
PCR Genotyping AT4G33280
Wild Type 25 Homo T-DNA 0 Hemi T-DNA 0
1 agarose 1 Hour 05/15/2008 105 Volts
15
Carrying Out More Genotyping
PCR Genotyping AT4G33280
1 agarose 05/29/08 1Hr. Run 105 Volts
16
Genotyping With Controls
PCR Genotyping of Plants 7, 17, 26, 33
1 Gel 05/30/08 1 Hr.Run 105 Volts
17
AT4G33280's Activity
RT-PCR AT4G33280
534-712 pm 1.5 Agarose 05/20/2008 105 Volts
Expected Product 254bp
18
(No Transcript)
19
Cloning the Upstream Region
Colony Digestion
pENTR Gene
Vector
Promoter Non-Recombinant 2580bp
262bp Recombinant 2842bp

1 Agarose, 1 Hour Run, 1 agarose, 105 Volts
20
Nomarski
21
THANKS TO

• Dr. Goldberg
• Anhthu Bui
• Bekah Charney
• Daisy Robinton
• Brandon Le
• Tomokazu Kawashima