Alfred and martha used the high setting the shear

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- Dump Bacteria with attached Phage into a Blender and turn it on. (Alfred and Martha used the “High” setting.) The Shear Forces in the Blender will strip the Bacteriophage “Ghosts” off the Surface of the Bacteria. - Pour the resultant Cocktail into Centrifuge Tubes and spin-down the Bacteria The Phage “Ghosts” will remain in the Supernatant (G= to swim above) 35 S 32 P Remains in Phage Sediments with “ghosts” in Supernatant Bacteria in the Pellet • Allow infected Bacteria in the Pellets enough Time for one Round of Phage Replication 35 S 32 P No Phage produced Phage produced containing 35 S containing 32 P The Waring Blender Experiment demonstrated that Phage DNA alone contains all Information required for making more Phage • And this explained what was going on during the Eclipse Period
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Bacteriophage Page 5 Bacteriophage T4 Replication • Attachment Tail Fibers make contact. The Baseplate is lowered onto the Cell Surface. The Outer Sheath Contracts and Phage DNA is inserted (injected) into the Cytoplasm. • Replication - T4 mRNA Synthesis (Transcription) begins Early Proteins are transcribed by unmodified E. coli RNA Polymerase within Seconds of Infection. Within Minutes increasingly T4-Midified (“Customized”) E. coli RNA Polymerase begins to transcribe Middle and Late Proteins. - T4 Protein Synthesis (Translation) T4 mRNA is obligingly translated by E. coli 70S Ribosomes within MInutes. - T4 DNA Synthesis (Replication) begins T4 DNA Synthesis involves a T4-Coded DNA Polymerase. The Synthesis of T4 DNA begins at T= 5 Minutes. • Assembly - Separate Sub-Assemblies are made for the Head and Tail. Head Sub-Assembly Tail Sub-Assembly Assisted Assembly of Procapsid Baseplate Assembly | | Mature Capsid (“Head”) Central Tube Polymerization | | DNA Inserted into Head Outer Sheath Polymerization \ / Final Assembly Head Joined to Tail | Tail Fibers Added | Bacteriophge T4 • Release - T4 encodes for the Enzyme Lysozyme • Lysozyme lyses the E. coli like a Piñata
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