Lecture11_NABPty2012 (1)

4 per base pair 5 3 minor major groove 34 10 bp per

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Unformatted text preview: s. In solution, there are ~10.5 base-pairs per turn. To find whether the two strands are parallel or antiparallel, Kornberg synthesized DNA enzymatically from labeled nucleoside triphosphates 1. Synthesize DNA from a mixture of ATP, GTP, CTP & TTP in which one of the four nucleotides is labeled with 32P in the α position (e.g., pppA). 2. Hydrolyze the DNA with a DNase that releases the 3’-mononucleotides. 3. Measure the amount of 32P transferred to various nucleotides. pppA + pppT + pppG + pppC DNA polymerase 5’ ... pGpApTpCpApGp ... 3’ 3’ ... pCpTpApGpTpCp ... 5’ ... Gp + Ap + Tp + Cp + Ap + Gp ... H2O DNase ... Cp + Tp + Ap + Gp + Tp + Cp ... 4. Repeat the experiment starting with the label in other nucleotides. 4. If the strands are antiparallel: A C = G T and A G = C where the arrows indicate the transfer pf the radioactive phospate/ What if the chains are parallel? High-resolution structures of small DNA molecules have been obtained by x-ray crystallography and NMR NMR NMR structure of a duplex DNA dodecamer in the DNA-binding site of an interferon promotor. J. R. Huth et al., Nature Struct. Biol. 4: 657 (1997). 2eze.pdb The two strands of the double helix separate reversibly at...
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This note was uploaded on 02/17/2012 for the course CHEM 212 taught by Professor Staff during the Fall '10 term at Rutgers.

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