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What would you like discussed in class, on any subject that has already come up?

Welcome to Molecular Biology Through Discovery Tuesday, 18 September 2012 DNA Structure / Sanger & Tuppy. General Questions. What would you like discussed in class, on any subject that has already come up?.

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What would you like discussed in class, on any subject that has already come up?

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  1. Welcome toMolecular Biology Through DiscoveryTuesday, 18 September 2012DNA Structure / Sanger & Tuppy

  2. General Questions What would you like discussed in class, on any subject that has already come up? The mentor list and clarification on who, what, when, etc. Who can we pick, how many can we choose from? What are we turning in exactly on Thursday???

  3. DNA Structure ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ... TAGGCCACTGCCCAACCCTCCATCATAAAACTTGGGCTTGGGAGGCAGAGCCTAACCTCTCTCACTCTAGACAGGTCTAAGATGATTGGGAACGAAATGAGCCGTCTCGACTTTTTCGCGAAGTGGCTAA ... #1: Hello! #2: Complete description of our civilization

  4. DNA Structure Biology Today and Tomorrow Starr, Evers, and Starr (2010)

  5. DNA Structure Biology: Understanding Life Alters (2000)

  6. DNA Structure E. How can the helical structure of DNA and internucleotide distance be discerned from Franklin and Gosling's x-ray photograph?

  7. DNA Structure E. How can the helical structure of DNA and internucleotide distance be discerned from Franklin and Gosling's x-ray photograph?

  8. DNA Structure It's necessary to be slightly underemployed if you are to do something significant. - Jim Watson

  9. NH2 N N N N O NH2 O N O O O O O O O O N NH NH N P P P P N - - - - O O O O O O O O N N O N O NH2 From the nucleotides shown above, construct a double-stranded DNA fragment with the sequence ACTG. You may: duplicate (Ctrl-d) horizontal flip (Alt-hgoh) vertical flip (Alt-hgov) and/or rotate (Alt-hgor) the nucleotides, but you may not change the relative positions of their atoms.

  10. DNA Directionality & Palindromes SQ10. If one strand of DNA had the sequence 5'-GGACT-3', what would be the sequence of the second strand?

  11. DNA Directionality & Palindromes I understand what a palindrome is in English but when it comes to DNA how come 5'-AGTTGA-3' isn't a palindrome when it's anti-parallel strand is 3'-TCAACT-5' which is also a palindrome.

  12. Backwards = forwards GCTATCG • DNA is double stranded ROTATOR TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA Palindromic Sequences What is it? What about with DNA?

  13. Backwards = forwards GCTATCG ROTATOR Palindromic Sequences What is it? What about with DNA? • DNA is double stranded • DNA is redundant TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

  14. Backwards = forwards GCTATCG ROTATOR TTAATGTGAGTTAGCTCACTCATT AATGAGTGAGCTAACTCACATTAA Palindromic Sequences What is it? What about with DNA? • DNA is double stranded • DNA is redundant • DNA has direction (read 5’->3’) 5’- -3’ 3’- -5’ TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

  15. TAT GGCATGCTAGCTTAAT TCATTAATTA AGTAACGTACGATCGG TAT DNA: cruciform RNA: stem/loop Palindromic Sequences Palindromic sequences as structural RNA 5’- -3’ 3’- -5’ TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

  16. UAU GGCAUGCUAGCUUAAU UCAUU tRNA DNA: cruciform RNA: stem/loop Palindromic Sequences Palindromic sequences as structural RNA 5’- -3’ 3’- -5’ TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

  17. Palindromic Sequences Palindromic sequences as protein binding sites why [are] palindromes… targeted by DNA-binding proteins why palindromes are targeted by DNA-binding proteins

  18. TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA recognizes GTGAGTT Palindromic Sequences Palindromic sequences as protein binding sites

  19. TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA Palindromic Sequences Palindromic sequences as protein binding sites

  20. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA

  21. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA

  22. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA

  23. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA recognizes GTGAGTT

  24. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA

  25. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA

  26. Palindromic Sequences Palindromic sequences as protein binding sites TTAATGTGAGTTAGCTCACTCATT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNN AATGAGTGAGCTAACTCACATTAA Palindromes: Serve as binding sites for dimeric protein

  27. RNA Polymerase Palindromic Sequences Palindromic sequences as protein binding sites gene 5’-GTA ..(8).. TACNNNNNNNNNNTANNNTNNNNNNNNNNNNNNNNNNNNNNNNNNNNATGNNNNNNNNNNNNNNNN3’-CAT ..(8).. ATGNNNNNNNNNNATNNNANNNNNNNNNNNNNNNNNNNNNNNNNNNNTACNNNNNNNNNNNNNNNN GTA ..(8).. TAC

  28. gene 5’-GTA ..(8).. TACNNNNNNNNNNTANNNTNNNNNNNNNNNNNNNNNNNNNNNNNNNNATGNNNNNNNNNNNNNNNN3’-CAT ..(8).. ATGNNNNNNNNNNATNNNANNNNNNNNNNNNNNNNNNNNNNNNNNNNTACNNNNNNNNNNNNNNNN Transcription factor RNA Polymerase Palindromic Sequences Palindromic sequences as protein binding sites RNA GTA ..(8).. TAC Is the promoter a beginning string of nucleotides for RNA,

  29. Sanger and Tuppy (1951) Phe-Val-Asp-Glu-His-Leu-Cys-Gly Thr-Pro-Lys-Ala Gly-Glu-Arg-Gly-Tyr-Leu-Val-Cys-Gly Ser-His-Leu-Val-Glu-Ala

  30. Sanger and Tuppy (1951)

  31. Sanger and Tuppy (1951) Insulin is a dimer, composed of one A chain (fraction A) and a B chain (fraction B). Sanger and Tuppy determined this experimentally by oxidizing insulin with performic acid. The sequence of fraction B was determined experimentally as being at least composed of phenylalanine, valine, aspartic acid, and glutamic acid, and well as threonine, proline, lysine, and alanine. This was done using a prepared sample of polypeptide fragments, which were separated through a process known as paper chromatography. A similar process was done for fraction A. From all the studies perform, the overall structure and cross linking between chains can be deduced. Cross-linking can occur on cysteine residues, because they contain sulfur atoms, capable of dimerizing with one another and forming stable covalent bonds. Overall, two phenylalanine and two glcyl chains were determined to be the subunits of this particular form of insulin.

  32. Sanger and Tuppy (1951) Now, how this was deduced was through a number of chromatography tests where different chemicals were used in order to split insulin at different bond points and then examine the fragments. In the first section we can see Phe.Val.Asp.Glu.His.Leu.CySO3H.Gly which has been determined by the following: B4β2, B1α2, B2γ8, B1α1, B1β8, B1γ1, B1α6, B1β13, B1γ4, B1β10, B1α5, B1γ7, B1β12, B4β1, B1γ6, B1β5. Where B1α1 refers to the test number and type, then spot number.

  33. Sanger and Tuppy (1951) 1. Cys-Gly B1a.1

  34. Sanger and Tuppy (1951) 1. Cys-Gly B1a.1

  35. Sanger and Tuppy (1951) 1. Cys-Gly B1a.12. Leu-Cys B1a.6

  36. Sanger and Tuppy (1951) 1. Cys-Gly B1a.12. Leu-Cys B1a.63. Leu-Cys-Gly #1 & #2

  37. Sanger and Tuppy (1951)

  38. Sanger and Tuppy (1951) 1. Cys-Gly B1a.12. Leu-Cys B1a.63. Leu-Cys-Gly #1 & #2

  39. Results vs Conclusions Kaleigh Jonathan Michael Colleen AbdulCailin Sue Kristen Bobby Supriya Tayab Shahroze Mandi Shaun Abdallah Celeste Neda Yordanos Me And go to CyanoBIKE

  40. Sanger and Tuppy (1951)

  41. Coming Attractions

  42. Benzer (1959)

  43. Goodbye fromMolecular Biology Through DiscoveryTuesday, 18 September 2012 ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ... G-O-O-D-B-Y-E

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