Heat Shock Factor Protein Family of Transcription Factors

1. Heat Shock Factor Protein Family of Transcription Factors By Brian Yu

2. So…What is the question? What Leads to the formation of:

3. 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.

5. Genotyping Gels: At5g62020 At4g11660

6. General Information about the two genes:

7. 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’ Exon Exon 5’ UTR 3’ UTR At4g11660 SALK predicted location (Wrong orientation) *not to scale

8. RT-PCR data and GeneChip data for At5g62020

9. Inflorescence RT PCR Results for At4g11660

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

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

12. Any Observable phenotypes in the seed? Mut At5g62020 Mut At4g11660 WT

13. Observable Phenotypic Variations in the seeds? Mut At5g62020 Mut At4g11660 WT

14. 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”