BCHM 218 Study Guide - Quiz Guide: Aaa Proteins, Endonuclease, Dna Clamp
7 Dec 2015
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Document Summary
Dna replication: simple way to observe activity of dna pol, measures incorporation of a radioactively or fluorescently labeled dntp into the high molecular weight polymer, dna. Detection of dna polymer: precipitation of the polymer with a strong acid or positive filter in low salt (unincorporated nucleotides or nucleosides do not precipitate, analyze radioactivity by geiger counter, scintillation counting or electrophoresis and audioradiography. Identification and isolation of dna pol activity from other proteins can be done by chromatography. Dna pol requires a template strand, primer strand, dntps and mg2+ Dna polymerases are not very conserved: structures/sequences quite different from each other, bacteria can make longer stretches of okazaki fragments, faster. Dna replication is semi-conservative: each daughter strand contains one parental and one newly synthesized strand. Replication is initiated at specific sequences: eukaryotes have many, bacteria not so many (even one works!) Replication is typically bidirectional: replication fork moves away from ori in both directions.
