2. Virus Replication Hsiao-Sheng Liu 劉校生 國立成功大學微生物暨免疫學研究所

3. The objectives: How to calculate virus titer? One step growth curve The general stages of viral replication cycle Attachment, penetration and release of the viruses

4. How to measure virus titer?

5. Baculovirus plaques Zones of clearing (plaques) are generated by infection of insect (Sf9} cells with individual baculovirus particles. Uninfected Sf9 cells surrounding the plaque are stained pink with neutral red.

6. 10 -6 10 -7 PFU: plaque forming unit 1 PFU = 1 plaque = 1 bacterial phage Count : plaque 數量 x 稀釋倍數 x 10/9 = PFU/ml Ex: 211 x 10 7 x 10/9 = 2.34 x 10 9 PFU/ml 211 plaques

7. What is one step growth curve?

8. 陰蔽期 潛伏期

10. What are the general stages in viral replication cycles?

11. General replication procedure

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14. Complicated replication procedure (e.g. Herpes virus)

15. How the viral replication cycle is known?

17. Attachment: receptor, tissue tropism

18. HA: Hemagglutinine

19. Trimmers of HA on influenza virus membrane

20. Coreceptor: CCR5 CRCX4

21. CCR5: macrophage-tropic HIV CXCR4: lymphocyte-tropic HIV

22. How does the virus enter the cell?

23. Endocytosis Fusion Pinocytosis (Viropexis)

24. Paramyxovirus fusion  Syncytia

25. Togavirus

26. How is the virus released from the cell?

27. Orthomyxovirus with M protein Herpes virus

28. Respiratory syncytial virus budding

29. Brooks et al, 1998