11.03.2011 - Lecture 17 November 3 2011 Gene Isolation and...

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Lecture 17 November 3, 2011 Gene Isolation and Manipulation (Ch. 10)
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Translation Initiation in Prokaryotes IF1, IF2, IF3
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Translation Initiation in Eukaryotes eIF4A,B, and G
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Translation Initiation Translation Elongation Translation Termination
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EF-Tu, EF-G Translation Elongation:
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Translation Initiation Translation Elongation Translation Termination
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Translation Termination: Stop codons: UGA, UAA, UAG Release factors RF1, RF2, RF3 RF1,RF2 Stop codons RFs in A site No peptide bond H2O in peptidyltransferase center Polypeptide release from tRNA
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Gene Cloning: Relevant Tools And Techniques
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Generating Recombinant DNA Molecules Restriction enzymes – cleave phosphodiester bonds at specific sequences 5’-GAATTC-3’ 3’-CTTAAG-5’ Eco RI : Restriction sites are typically palindromic “sticky” ends
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1. Load DNA in wells 2. DNA migrates toward “+” pole. 3. Stain gel with ethidium bromide
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11.03.2011 - Lecture 17 November 3 2011 Gene Isolation and...

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