do at4g13640 and at3g24120 play a significant role in seed development for arabidopsis thaliana ?
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Do AT4G13640 and AT3G24120 Play a Significant Role in Seed Development for Arabidopsis Thaliana ?. By Jordan Fischer June 8, 2006 Professor Goldberg hc70al. Transcription factors are proteins involved in the regulation of gene expression. - PowerPoint PPT PresentationTRANSCRIPT
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Do AT4G13640 and AT3G24120 Play a Significant Role in Seed Development
for Arabidopsis Thaliana?
By Jordan Fischer
June 8, 2006
Professor Goldberg
hc70al
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What Is a Transcription Factor?
• Transcription factors are proteins involved in the regulation of gene expression.
• Bind to the promoter region upstream of gene and either facilitate or inhibit transcription.
• Control and regulate gene expression.
• Activated by:– Physiological stimuli.– Therapeutically.– Pathological stimuli.
Examples of Transcription Factors
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What Is the Significance of the G2-like Transcription Factor Family?
• Recently categorized GARP super family of transcription factors defined by G2 in maize; the Arabidopsis RESPONSE REGULATOR-B proteins; and the PHOSPHATE STARVATION RESPONSE1 protein of Chlamydomonas.
• G2 function is specifically committed to the differentiation of bundle sheath cell chloroplasts in C4 leaf blades.
• Act as transcriptional regulators of cell-type differentiation processes.
• Likely that GLK proteins act as transcriptional regulators of chloroplast development.
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What is the Structure of Gene AT4G13640?
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What is the Structure of Gene AT3G24120?
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What Other Information is Important Pertaining to These Genes?
AT4G13640 AT3G24120
Protein Type MYB MYB
Size (bp) 2039 2436
Size (aa) 292 295
# Exons 6 6
# Introns 5 5
Most Active 24-Hr. Seed Ovule and 24-Hr. Seed
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According to Gene Chip, Where are These Genes Active in the Arabidopsis Plant?
Gene AT4G13640Gene AT3G24120
Control (Gene AT1G74840)
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Are the Results from RT-PCR Consistent with
Those of Gene Chip? AT4G13640 AT3G24120
Leaf +
RT
Leaf -R
T
Silique +
RT
Silique -R
T
Leaf +
RT
Leaf -R
T
Silique +
RT
Silique -R
T
(+) C
ontrol
(+) C
ontrol
Yes, according to Gene Chip and RT-PCR both genes are active in the leaf and silique of the Arabidopsis plant.
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Did My Experiments Agree With Salk’s Regarding the T-DNA Insert Location?
AT4G13640: YES
AT3G24120: NO
5’UTR
5’UTR
3’UTR
3’UTR
Exon 1 Exon 2 Exon 3 Exon 4 Exon 5 Exon 6
Exon 1 Exon 2 Exon 3 Exon 4 Exon 5 Exon 6
LbB1
LbB1
Rev.Fw.
Rev.Fw.
LbB1
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What Genotyping Data Was Collected For Gene AT4G13640?
1st Batch 2nd Batch
1 Kb L
adderC
ontrol (-)C
ontrol (+)
Plant #18Plant #17Plant #16Plant #15Plant #14Plant #13Plant #12Plant #11Plant #10Plant #9Plant #8Plant # 71 K
b Ladder
Control (-)
Control (+
)P
lant #6P
lant #5P
lant #4P
lant #3P
lant #2P
lant #11 K
b Ladder
Wild Type Band
Mutant Type Band
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What Genotyping Data Was Collected For Gene AT3G24120?
RV & LBb1 FW & LBb1 FW & RV LBb1 Cont.
Control (-)
Control (+
)P
lant #17P
lant #16P
lant #15P
lant #14P
lant #13P
lant #12P
lant #111 K
b Ladder
Control (-)
Control (+
)P
lant #17P
lant #16P
lant #15P
lant #14P
lant #13P
lant #12P
lant #11
1 Kb L
adderL
Bb1 C
ontrolC
ontrol (-)C
ontrol (+)
Plant #17
Plant #16
Plant #15
Plant #14
Plant #13
Plant #12
Plant #11
Wild Type Band
Mutant Type Band
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What is the Summary of Genotyping Results for Genes AT4G13640 and AT3G24120?
AT4G13640 AT3G24120
Wild/Wild
Mutant/Mutant
Heterozygous
TOTAL
5 8
2 5
50
7 18
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How Did the Phenotypes of Homozygous Mutant and Wild Type Plants Compare?
Mutant Wild Type
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How is Promoter Cloning Used in this Experiment?
Colony # 4
Colony #3
Colony #1
1 Kb L
adder
(-) Control
(+) C
ontrolA
mp. R
egion1 K
b Ladder
(-) Control
(+) C
ontrolA
mp. R
egion1 K
b Ladder
Colony # 4
Colony #3
Colony #2
Colony #1
1 Kb L
adderAT4G13640
AT3G24120
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What Conclusions Can Be Made Based on these Experiments?
• Genes AT4G13640 and AT3G24120 are not active in seed development to the extent where a knock out of either gene will be lethal to seed development.
• A knockout of these genes causes no observable phenotypical differences compared to the wild type.
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What Further Experiments Should be Done on Genes AT4G13640 and AT3G24120?
• Use GFP as a marker to help to determine where in the plant the promoter regions activate transcription.
• Cross homozygous mutant plants from both knock out lines and determine if a complete knock out of both genes is fatal to seed development or if this creates any phenotypic differences.
• Use Nomarksi microscope to check for phenotypical differences in seed development.
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