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Unformatted text preview: Grow bacteria in liquid broth medium Add phage to bacterial cells Enough phage to infect all the bacterial cells (e.g., an MOI of 5-10 = 5-10 phage per cell) Allow attachment, then remove unadsorbed phage from cells and add fresh medium Remove samples at regular intervals and determine the number of phage present (extracellular and intracellular) by a plaque assay Plot total # of phage (or PFU) per milliliter of sample verses time One-Step Growth Curve of a Phage One-Step Growth Curve of a Phage Eclipse (no infectious virion inside or outside bacteria) Attachment to host Transfer of genome Synthesis of proteins and genome Intracellular maturation assembly of whole virion within cell + Eclipse = Latent period Rise Release of virion from cell Plateau All cells lysed and all virion liberated Phage typically reaches a plateau in less than an hour Modified Figure 6.28 Burst size = # of virions produced per infected cell...
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- Spring '08