Transcript Slide 1

Heat Shock Factor Protein
Family of Transcription Factors
By Brian Yu
So…What is the question?
What
Leads to the formation of:
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.
Genotyping Gels:
At5g62020
At4g11660
General Information about the two
genes:
At5g62020
At4g11660
Gene Size
(bp)
1555 bp
2076 bp
# amino
acids
300 aa
378 aa
#TDNA
inserts
(actual)
#TDNA
inserts
(predicted)
2
2
1
1
Graphical Interpretation of tDNA
insert area
TDNA insert 5’->3’
TDNA 3’->5’
At5g62020
5’ UTR
Exon
Exon
3’ UTR
SALK predicted location
(wrong orientation)
TDNA insert 5’ -> 3’
TDNA insert 3’ -> 5’
5’ UTR
Exon
Exon
3’ UTR
At4g11660
SALK predicted location (Wrong orientation)
*not to scale
RT-PCR data and GeneChip data
for At5g62020
RT PCR Results for At4g11660
Promoter Region Experiment:
EcoR1 Digest for pAt4g11660
EcoR1 Digest for pAt5g62020
Orientation of the insert in Topo
Plasmid (At5g62020)
5’
SP6 Site
3kB
Promoter Region
Topo Plasmid
3’
T7 Site
Any Observable phenotypes in the
seed?
Mut
At5g62020
Mut
At4g11660
WT
Observable Phenotypic Variations
in the seeds?
Mut
At5g62020
Mut
At4g11660
WT
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”