MBIO 3410 Lecture Notes - Lecture 1: Klenow Fragment, Exonuclease, Dna Replication
24 Dec 2015
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In e. coli, other dna polymerases: dna pol iv and v. Klenow fragment is dna polymerase i from e. coli with 5"3" (cid:1) exonuclease activity removed. So it doesn"t degrade any dna you are trying to make. Phage polymerases are more stable, have higher processivity, single (cid:1) subunit, and no 5"3" exonuclease activity. Topoisomerase nicks one or both strands of dna to reduce the stress of unwinding and then re-ligates them. Origin of replication: dna sequence recognized by helicase (or (cid:1) (cid:1) (cid:1) (cid:1) (cid:1) gyrase) and dna is unwound. Primer is laid out by primase (rna primer on dna template) Ssbps bind single strands to prevent the two strands from re- Dna polymerase units attached to dna via beta clamp. Dna polymerase iii complex has two polymerase catalytic annealing components. Clamp loader: protein that will guide beta clamp into new positions and reconnect to dna polymerase. Replisome: molecular machinery that carries out dna replication.
