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My Summer

My Summer. ~ 在風裡的男人 ~ 891607 林明興. 中研院 分生所 Dr. Huey-Nan Wu 吳惠南. 研究方向. D 型肝炎病毒抗原核酸監護子活性的研究 登革熱病毒複製機制的研究. C prM E NS1 NS2 NS3 NS4 NS5. Introduction (I). Dengue Virus. S.S. RNA. NS3 N-terminal Helicase NS3 C-terminal Protease

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My Summer

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  1. My Summer ~在風裡的男人~ 891607 林明興

  2. 中研院 分生所 Dr. Huey-Nan Wu 吳惠南

  3. 研究方向 • D型肝炎病毒抗原核酸監護子活性的研究 • 登革熱病毒複製機制的研究

  4. C prM E NS1 NS2 NS3 NS4 NS5 Introduction (I) Dengue Virus S.S. RNA NS3 N-terminal Helicase NS3 C-terminal Protease NS5 N-terminal RdRp NS5 C-terminal Methylase Structure protein Non-structure protein

  5. RNA cDNA Clone Introduction (II) Expression Purification

  6. My Project Clone, expression and purification for DENGUE virus type II & III NS5 N-terminal domain Protein

  7. Clone for Dengue NS5-N • Amplify cDNA • Recombine to vector, pET21b • Transform to competent cell, JM101 • Checking with restriction enzyme

  8. Expression • Recombinant DNA transform to competent cell, BL21DE3 • Cell culture • Adding IPTG to induce protein expression • SDS-PAGE analysis

  9. DNII5N-11 positive control

  10. Protein purification • Reagent preparation • Cell extract preparation • Resin preparation (charge with Ni+) • Cell extract flow through column • Eluting the target protein

  11. Small scale purification – batch method • Purifying with various salt concentration • Eluting without denature reagent, urea • Eluting with lower concentration imidazole

  12. Various NaCl concentration

  13. Urea & Imidazole

  14. Imidazole

  15. Large-amount purification – column chromatography

  16. Large-amount purification – column chromatography

  17. Some memories

  18. We can see more!

  19. 我講完了 請鼓掌

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