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DNA synthesis - Q7-15 rely on the following info: Ó-PO₂H NH₂ N While nearly all organisms on Earth use a DNA code of ATCG, a particular class of bacteriophages (viruses that infect bacteria), was found to use ZTCG, where Z is an 2'aminoadenine, a modified form of adenine. dZTP basepairs with thymine in the phage DNA, forming 3 hydrogen bonds. NH₂ HO 7) Draw an AT basepair and a ZT basepair, including the riboses and a phosphoryl group. (16 points) 8) Draw an arrow to the additional hydrogen bond in ZT. (4 points) 9) Draw an arrow to a glycosidic bond. (4 points) 10) Draw arcs indicating the major and minor grooves for both AT and ZT basepairs. (4 points) PO3H2 O-PO₂H
Q 14-15 refer to the follosing information (and also refer to questions 7-13). A ATP 1 5'-FAM-CCCCTTATTAGCGTTTGCCC To test the activity of each polymerase with dATP and dZTP, the researchers performed 3-GGAATAATCGCAAACGGGTTT TTTTTTTT-5 dZTP 25-FAM-CCCCTTATTAGCGTTTGCCC a primer extension 3-GGAATAATCGCAAACGGOTT assay with each purified polymerase where a fluorescently labeled primer (FAM is a fluorescent label) was annealed to a DNA template such that extension required polymerization of either dATP or dZTP. DNA polymerase KM (JM) Keat (5¹) dZTP dATP dZTP dATP 13.96 E. coli Klenow fragment Vibrio phage PhiVCS 0.81 0.14 0.70 0.17 0.86 0.10 0.02 4.50 1.38 45.00 Acinetobacter phage SH-Ab 15497 1.300.36 21.40 3.40 16.47 1.99 20.61 * 1.52 0.42 0.14 0.21 0.05 0.29 0.07 0.16 0.01 14) Given the data on keat and Km above, which of the following gels for the primer extension assay corresponds the extension with the E. coli Klenow fragment and which to the Acinetobacter phage polymerase? Note that the first two lanes show the full length product and initial primer, and the lanes labeled 1 and 2 correspond to the 6 situations presented above (1 = extension with dATP, 2-extension with dZTP). Explain your answer with a discussion of the Km and keat values provided above. (15 points) Gel A-polymerase 1 Gel B-polymerase 2 12 Full length Full length Prime Primer 15) The primer extension assays also contained the following components. Discuss why they are important for the reaction: a) 20 mM Tris-HCl (pH 7.5) (5 points) b) 5 mM MgCl₂ (5 points)