Concatemers large linear dna oligomers in t7 formed

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Unformatted text preview: 00kDa) •  T7 RNA polymerase transcribes Class II and Class III promoters •  T7 RNA polymerase takes over from host RNA polymerase in transporGng DNA into cell T7 DNA replication •  BidirecGonal •  Resembles E.coli DNA synthesis •  ConGnuous leading strand •  DisconGnuous lagging strand •  T7 encodes all it own DNA replicaGon proteins •  Unique Origin 5.9kb from let end of genome •  Prevents RNA polymerase – DNA replicaGon fork collisions •  DNA replicaGon is iniGated by T7 RNA polymerase •  Large concatemers of T7 genomes are formed via 150bp terminal repeats T7 DNA replication T7 Primase/Helicase 3’ T7 DNA polymerase 3’ 3’ 5’ 3’ T7 RNA polymerase 3’ Concatemer Formation •  Problem •  DNA polymerase needs a primer to iniGate DNA synthesis •  Thus, the RNA primer at 3’ end of template DNA cannot be removed or replaced with DNA •  This would result in the loss of sequence •  T7 soluGon •  Concatemers Concatemers •  Concatemers •  Large, linear DNA oligomers •  In T7 formed via the 150bp terminal repeats •  Concatemers are cleaved i...
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