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What I wanted to do this summer

What I wanted to do this summer Evaluate inducible inactivation system designed to regulate mitochondrial gene expression Examine unintended consequences of TAP-tagging. What I really did this summer. Targets for protein engineering. Pilot studies. = no TAP-tag. FY631

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What I wanted to do this summer

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  1. What I wanted to do this summer Evaluate inducible inactivation system designed to regulate mitochondrial gene expression Examine unintended consequences of TAP-tagging What I really did this summer

  2. Targets for protein engineering

  3. Pilot studies = no TAP-tag FY631 MAT(A) his4-917, lys2-173R2, leu21, ura3-52, trp163 = SPT3-TAP = SPT7-TAP = SPT10-TAP FY2305 MAT(A) his3200, lys2-128, leu21, ura3-52, trp163

  4. HIS3 as expected SPT10: most affected genes, pt 1 higher - TAP

  5. SPT10: most affected genes, pt 1 higher - TAP LYS2 No TAP lys2-173R2 vs +TAP lys2-128

  6. F0-ATP synthase subunit 9 encoded on the mitochondrial genome Ask: OligomycinR? SPT10: most affected genes, pt 1 higher - TAP OLI1 again! Also in SPT3-TAP and SPT7-TAP

  7. SPT10: most affected genes, pt 1 higher - TAP mitochondrial genes

  8. SPT10: most affected genes, pt 2 higher + TAP SPT10 Increased expression of SPT10-TAP vs SPT10

  9. SPT10: most affected genes, pt 2 higher + TAP PHO5 and PHO11

  10. SPT10: most affected genes, pt 2 higher + TAP ADH2 (alcohol dehydrogenase) and ADY2 (Acetate transporter required for normal sporulation; phosphorylated in mitochondria) Also in SPT3-TAP and SPT7-TAP

  11. End of summer ppt

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