Chemistry Chapter 26
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1 Chemistry 2100 Chapter 26
2 The Central Dogma! The central dogma of molecular biology: Information contained in DNA molecules is expressed in the structure of proteins. Gene expression is the turning on or activation of a gene. 2!
3 transcription
4 transcription
5 transcription
6 Transcription! The architecture of yeast RNA polymerase II. Transcription of DNA (helical structure) into RNA (red) is shown. The template strand of DNA (blue) and the coding strand (green) and also shown. 6!
7 Transcription! Organization and transcription of a split eukaryote gene. 7!
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12 Why detect Transcrip6on Factor targets? Transcrip6on factors are medically relevant ~10% of human genes Crucial roles in development and cell life cycle Misregula6on and muta6on cause disease Cri6cally, most cancers involve TF overac6vity Darnell, Nature Reviews Cancer 2, 740 (2002)
13 Tradi6onal methods for Transcrip6on Factor detec6on Expression Microarrays Western Blots Gel Shift Assays The challenge: Most of these methods are indirect, slow (hours), or can t differentiate active and inactive protein.
14 Marvin J S et al. PNAS 1997;94: Bio- mimicry is a powerful mo6va6on Velcro: inspired by burrs Conformation Switching Probes
15 Op6cal Conforma6on Switching TF Switch Sensors
![Ra6onally Tuning TF Sensors KS = [ ] [ ] % switches open = [ ] [ ] KD = [ ] KS](/docs-images/80/81017540/images/16-0.jpg "[target] KD (1+ KS) + KS [target] % switches open KS = 10 KS = 1 KS = 0.")
16 Ra6onally Tuning TF Sensors KS = [ ] [ ] % switches open = [ ] [ ] KD = [ ] KS [target] KD (1+ KS) + KS [target] % switches open KS = 10 KS = 1 KS = 0.1 Target [M]
17 TF Beacon Actual Performance
18 Quan6ta6ve Detec6on in 4 easy steps
19
20 initiation
21 initiation
22 initiation
23 elongation
24 elongation
25 elongation
26 elongation
27 elongation
28 elongation
29 elongation
30 mrna trna (5') A U G U U U G U C A G U..... U A C A A A C A G U C A peptide Met Phe Val Ser....
31 N 2 O O N ONO N N O N 2 O O N O N C U U A
32 Pyrimidine Dimers from UV
33 The Genetic Code! 33!
34 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
35 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
36 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
37 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
38 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
39 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
40 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr
41 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr COO COO GAA GAG 2 N GUA GUG 2 N C 2 C 2 COO Glu C(C 3 ) 2 Val
42 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr COO COO GAA GAG 2 N GUA GUG 2 N C 2 C 2 COO Glu C(C 3 ) 2 Val
43 UCU UCC UCA UCG 2 N COO C 2 O Ser ACU ACC ACA ACG 2 N COO O C 2 O Thr COO COO GAA GAG 2 N GUA GUG 2 N C 2 C 2 COO Glu C(C 3 ) 2 Val
44 DNA mrna (5') (3') (5') A T G T T T G T C A G T..... T A C A A A C A G T C A..... A U G U U U G U C A G U..... peptide Met Phe Val Ser....
45 DNA mrna (5') (3') (5') A T G T T T G T C A G T..... T A C A A A C A G T C A..... A U G U U U G U C A G U..... peptide Met Phe Val Ser....
46 DNA mrna (5') (3') (5') A T G T T T G T C A G T..... T A C A A A C A G T C A..... A U G U U U G U C A G U..... peptide Met Phe Val Ser....
47 DNA mrna (5') (3') (5') A T G T T T G T C A G T..... T A C A A A C A G T C A..... A U G U U U G U C A G U..... peptide Met Phe Val Ser....
48 mrna (5') Muta6ons A U G U U U G U C A G U..... peptide Met Phe Val Ser.... mrna (5') A U G U U U G A C A G U..... peptide Met Phe Asp Ser....
49 point substitution mutation mrna (5') A U G U U U G U C A G U A.... peptide Met Phe Val Ser.... mrna (5') A U G U U U G A C A G U..... peptide Met Phe Asp Ser....
50 point substitution mutation mrna (5') A U G U U U G U C A G U A.... peptide Met Phe Val Ser.... mrna (5') A U G U U U G A C A G U A.... peptide Met Phe Asp Ser....
51 point substitution mutation mrna (5') A U G U U U G U C A G U A.... peptide Met Phe Val Ser.... mrna peptide (5') A U G U U U G A C A G U A.... Met Phe Asp Ser....
52 frameshift mutation mrna (5') A U G U U U G U C A G U A.... peptide Met Phe Val Ser.... mrna peptide (5') A U G U U U G U C A G U A.... Met Phe Val Val....
53 frameshift mutation mrna (5') A U G U U U G U C A G U A.... peptide Met Phe Val Ser.... mrna peptide (5') A U G U U U G U C A G U A.... Met Phe Val Val....
54 frameshift mutation mrna (5') A U G U U U G U C A G U A.... peptide Met Phe Val Ser.... mrna peptide (5') A U G U U U G U C A G U A.... Met Phe Val Val....
55 PCR Polymerase Chain Reaction
56
57 DNA Fingerprin6ng
58 DNA Sequencing
59 DNA Sequencing
60 Shotgun Sequencing
61 Electrochemical Sequencing
62
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64 Recombinant DNA
65
66 Gene Therapy
Objective: You will be able to justify the claim that organisms share many conserved core processes and features.
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