Heat Shock Factor Protein Family of Transcription Factors

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: At 5 g 62020 At 4 g 11660

General Information about the two genes: At 5 g 62020 At 4 g 11660 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 t. DNA insert area TDNA 3’->5’ 5’ UTR TDNA insert 5’->3’ At 5 g 62020 Exon 3’ UTR SALK predicted location (wrong orientation) TDNA insert 5’ -> 3’ TDNA insert 3’ -> 5’ 5’ UTR Exon 3’ UTR At 4 g 11660 SALK predicted location (Wrong orientation) *not to scale

RT-PCR data and Gene. Chip data for At 5 g 62020

e e RT PCR Results for At 4 g 11660

Promoter Region Experiment: Eco. R 1 Digest for p. At 4 g 11660 Eco. R 1 Digest for p. At 5 g 62020

Orientation of the insert in Topo Plasmid (At 5 g 62020) 5’ SP 6 Site 3 k. B Promoter Region Topo Plasmid 3’ T 7 Site

Any Observable phenotypes in the seed? Mut At 5 g 62020 Mut At 4 g 11660 WT

Observable Phenotypic Variations in the seeds? Mut At 5 g 62020 Mut At 4 g 11660 WT

The future of the experiment • Sequence the plasmids for the experiment on At 4 g 11660. • Use the plasmids in further experiments with Green Fluorescence Protein to determine where in the promoter area is the “functional code”