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Thermus AQuaticus DNA polymerase SEQUENCE: MRGMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGFAKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPED FPRQLALIKELVDLLGLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLITPAWLWEKYGLRPDQWADYRALTGDESDNLPG VKGIGEKTARKLLEEWGSLEALLKNLDRLKPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLHEFGLLESPKALEEAPW PPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWT EEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAI GKTEKTGKRSTSAAVLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLCCCDPNLQNIPVRTPLGQRIRRGFIAEEGWLLVAL DYSQIELRVLAHLSGDENLIRVFQEGRDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERYFQSFPKVRAWIEKTLE EGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGV YPLAVPLEVEVGIGEDWLSAKE by Dave Phillis Spring 2010
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Dna Replication
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PCR Reaction
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Helices and Beta Barrels and side chains
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DNA and “O” helix
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Magnesium metal
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Magnesium Interactions
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ALL ZINC
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Hydrogen Bonding CTP
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HYDROPHOBIC INTERACTIONS
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ALL HYDRO AND H-BOND
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ALL CTP
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Sequence Alignment
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Literature Cited 1., F.; Stoffel, S.; Saiki, R.; Myambo, K.; Drummond, R.; Gelfand, D., (1989) Isolation, Characterization, and Expression in Escherichia coli of the DNA Polymerase Gene from Thermus aquaticus. Journal of Biological Chemistry. 264:6427-6437. 2. Patel, P.; Kawate, H.; Adman, E.; Loeb, L., (2000) A single highly mutable catalytic site amino acid is critical for DNA polymerase fidelity. Journal of Biological Chemistry. Nov 7. 3. Steitz, T. (1999) DNA Polymerases: Structural Diversity and common Mechanisms. Journal of Biological Chemistry. 274: 17395-17398. 4. Urs, U.; Ramachandran, M.; Krishna Murthy, H., (1999) Structure of Taq polymerase shows a new orientation for the structure-specific nuclease domain. Acta Cryst. D55:1971-1977. 5. Villbrant, B. ; Sobek, H. ; Frey, B.; Schomburg, D., (2000) Domain exchange: chimeras of thermus aquaticus DNA polymerase, escherichia DNA polymerase I and thermotoga neapolitana DNA polymerase. Protein Engineering. 9:645-54. 6. Kim, Y.; Eom, S.; Wang, J.; Lee, D.; Suh, S.; Steitz, T., (1995) Crystal structure of Thermus aquaticus DNA polymerase Nature 376, 612-616 (17 August 1995) 7. Chien A, Edgar DB, Trela JM (1976). "Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus". J. Bact. 127 (3): 1550–7. 8. Saiki, RK; et al. (1988). "Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.". Science 239 (4839): 487–91. 9. Saiki, RK; et al. (1985). "Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia" Science 230 (4732): 1350–4 10. Lawyer FC, et al. (1993). "High-level expression, purification, and enzymatic characterization of full-length Thermus aquaticus DNA polymerase...". PCR Methods Appl. 2 (4): 275–87. 11. Tindall KR and Kunkel TA (1988). "Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase". Biochemistry 27 (16): 6008–13
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