Arabidopsis Thaliana Gene

AT5G58610

Nancy Gallardo

HC70AL

What Protein Does AT5G58610 Code for?

• Regulates gene expression

• 1065 amino acids

PHD-Type Zinc Finger Transcription Factor

What's Around Gene AT5G58610 ?

AT5G8610 AT5G58620AT5G58600

AT5G58600

3' 5' 3' 5' 5' 3'

5' 3' 5' 3' 3' 5'

1045bp 2286bp

23703997-23708279

AT5G58620

5'

5'3'

3'

What is the Structure of AT5G58610 ?

Exon Exon Exon Exon Exon Exon Exon Exon

3'

5'

5'

3'

Chromosome 5

4283 base pairs

Reverse Orientation

9 Exons and 8 Introns

Exon

Expected T-DNA InsertionSalk_041367

Intron 2 Intron 1

T-DNA

546bp 362bp

Exon 1 Fw- 20bp

Rv- 20bp

LBb1-150bpExpected Sizes:WT= 908bpT-DNA= 696bp

3' 5'

What Were the Genotyping Results ?

PCR Genotyping AT5G58610

WT= 908 T-DNA=696

700bp

1% agarose, 105 Volts, 04/15/2008, 1.5Hour Run

Do these results agree with the RT-PCR results?

Where is AT5G58610 Active?

RT-PCR AT5G58610

1% Agarose 1 Hour Run 70 Volts 04/24/2008

Arabidopsis Thaliana Gene

AT4G33280

What Protein Does My Gene Code for?

B3-Domain Transcription Factor Families

Regulate gene expression 3 families in Arabidopsis Thaliana 337 amino acids

What's Around Gene AT4G33280 ?

AT4G33280 AT4G33290AT4G33270

3' 3' 5'

5' 5' 3'

561bp 217bp

16047358-16049359

5'

3' 5'

3'

What is the Structure of AT4G33280?

Exon Exon Exon Exon Exon Intron Intron Intron Intron

UTR UTR

•Chromosome 4

•2002 Base pairs

• Reverse Orientation

•5 Exons, 4 Introns, 2 UTRs

3'

5'

5'

3'

Expected T-DNA Insertion

Exon 5

AT4G32270 Intron 4

T-DNA

260Bp 142bp

LBb1-150bp

Fw-23bp

Rv-20bp

3'

Salk_072801

Expected Sizes:WT= 402bpT-DNA= 292bp

5'

What Were the Plants' Genotypes ?

Expected T-DNA=292bp Expected Wild Type=402bp

PCR Genotyping AT4G33280

Wild Type= 25

Homo T-DNA= 0

Hemi T-DNA= 0

1% agarose1 Hour 05/15/2008105 Volts

Carrying Out More Genotyping

PCR Genotyping AT4G33280

1% agarose05/29/081Hr. Run105 Volts

Genotyping With Controls

PCR Genotyping of Plants: 7, 17, 26, 33

1% Gel 05/30/08 1 Hr.Run 105 Volts

AT4G33280's Activity

RT-PCR AT4G33280

Expected Product= 254bp

5:34-7:12 pm1.5% Agarose05/20/2008105 Volts

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

Nomarski

THANKS TO:

Dr. Goldberg

Anhthu Bui

Bekah Charney

Daisy Robinton

Brandon Le

Tomokazu Kawashima