Heat Shock Factor Protein Family Heat Shock Factor Protein Family of Transcription Factorsof Transcription Factors

By Brian YuBy 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)

2 2

#TDNA inserts

(predicted)

1 1

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)

RT-PCR data and GeneChip data for At5g62020

RT PCR Results for At4g11660

Promoter Region Experiment:

EcoR1 Digest for pAt5g62020

EcoR1 Digest for pAt4g11660

Orientation of the insert in Topo Plasmid (At5g62020)

Topo Plasmid

Promoter RegionSP6 Site

T7 Site5’ 3kB 3’

Any Observable phenotypes in the seed?

WT

Mut

At5g62020

Mut

At4g11660

Observable Phenotypic Variations in the seeds?

WT

Mut

At5g62020

Mut

At4g11660

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”