Heat Shock Factor Protein Family of Transcription Factors
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Transcript of Heat Shock Factor Protein Family of Transcription Factors
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Heat Shock Factor Protein Family Heat Shock Factor Protein Family of Transcription Factorsof Transcription Factors
By Brian YuBy Brian Yu
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So…What is the question?
What
Leads to the formation of:
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What is a Heat Shock Factor Protein?
• It is a group of proteins that are formed when in the presence of heat (also seen when there is some stress to the organism)
• Found in all biological levels.
• Generally, these proteins serve as a way to help other proteins maintain their shape as heat will tend to denature the protein.
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Genotyping Gels: At5g62020 At4g11660
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General Information about the two genes:
At5g62020 At4g11660
Gene Size (bp)
1555 bp 2076 bp
# amino acids
300 aa 378 aa
#TDNA inserts (actual)
2 2
#TDNA inserts
(predicted)
1 1
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Graphical Interpretation of tDNA insert area
5’ UTR Exon Exon 3’ UTR
At5g62020
5’ UTR
At4g11660
Exon Exon 3’ UTR
TDNA insert 5’->3’TDNA 3’->5’
TDNA insert 3’ -> 5’TDNA insert 5’ -> 3’
*not to scale
SALK predicted location (Wrong orientation)
SALK predicted location (wrong orientation)
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RT-PCR data and GeneChip data for At5g62020
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RT PCR Results for At4g11660
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Promoter Region Experiment:
EcoR1 Digest for pAt5g62020
EcoR1 Digest for pAt4g11660
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Orientation of the insert in Topo Plasmid (At5g62020)
Topo Plasmid
Promoter RegionSP6 Site
T7 Site5’ 3kB 3’
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Any Observable phenotypes in the seed?
WT
Mut
At5g62020
Mut
At4g11660
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Observable Phenotypic Variations in the seeds?
WT
Mut
At5g62020
Mut
At4g11660
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The future of the experiment
• Sequence the plasmids for the experiment on At4g11660.
• Use the plasmids in further experiments with Green Fluorescence Protein to determine where in the promoter area is the “functional code”