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Erwin R. Schmidt
Institut für MolekulargenetikVorlesung #3
04. 05. 2015
Thema Gentechnologie
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Plasmide als Vektoren
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Blau-weiß-Selektion mit pUC-Vektoren
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Vektor zur Klonierung von PCR-Produkten
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Eigenschaften einiger PCR-Polymerasen
Table 1. Comparison of PCR Product Properties for Some Thermostable DNA Polymerases.
Taq/ VentR®/ Deep Characteristic AmpliTaq® Tfl Tth (Tli) VentR® Pfu Pwo
>95% >95% Resulting DNA ends 3´ A 3´ A 3´ A Blunt Blunt Blunt
5´®3´ exonuclease Activity Yes Yes Yes No No No No
3´®5´ exonucleaseactivity No No No Yes Yes Yes Yes
N.A.: not available
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Für die Expression fremder Gene gibt es spezielle Expressionsvektoren mit
Signalsequenzen für die korrekte Transkription und Translation in Bakterien
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Plasmidvektoren, zur Expression von Fremdgenen
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Expression von Fusionsproteinen mit sog. „His-tag“
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„Gateway“-Klonierungssystem
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„Gateway“-Klonierungssystem
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„Gateway“-Klonierungssystem
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„Gateway“-Klonierungssystem
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Replikation M13-Bakteriophage
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United States Patent 6803230 Inventors:
Bowdish, Katherine S. (Del Mar, CA) Fredrickson, Shana (Solana Beach, CA)
Wild, Martha (Solana Beach, CA)
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Substitutionsvektoren
Linker Arm „Stuffer“ Rechter Arm
Integrat
Ligation