Heat Shock Factor Protein Family of Transcription Factors By Brian Yu

So…What is the question? 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 #TDNA inserts (predicted) 1

Graphical Interpretation of tDNA insert area At5g62020 5’ UTR Exon Exon 3’ UTR SALK predicted location (wrong orientation) TDNA insert 5’ -> 3’ TDNA insert 3’ -> 5’ Exon Exon 5’ UTR 3’ UTR At4g11660 SALK predicted location (Wrong orientation) *not to scale

RT-PCR data and GeneChip data for At5g62020

Inflorescence RT PCR Results for At4g11660

Promoter Region Experiment: EcoR1 Digest for pAt4g11660 EcoR1 Digest for pAt5g62020

Orientation of the insert in Topo Plasmid (At5g62020) T7 Site 5’ 3kB 3’ SP6 Site Promoter Region Topo Plasmid

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”