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Identifizierung und Charakterisierung des HSP70 Makronukleus-Gens aus Euplotes crassus

Identifizierung und Charakterisierung des HSP70 Makronukleus-Gens aus Euplotes crassus

          
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About the Book

Inhaltsangabe: Zusammenfassung: Bei dem hypotrichen Ciliaten E.crassus wurde das Makronukleus-Chromosom, welches das HSP7O-Gen trägt, identifiziert. Für die Identifikation eines internen Gen-Fragmentes wurde die Methode der heterologen PCR angewendet. Die 5'- und 3'-Enden des Gens konnten mit einem telomerspezifischen Primer und mit homologen Primern, die aus dem internen PCR-Fragment abgeleitet wurden, amplifiziert werden. Nach Sequenzierung der PCR-Fragmente und einem Vergleich der abgeleiteten Aminosäuresequenzen mit Aminosäuresequenzen aus weiteren Organismen wurde eine Homologie von bis zu 95 % festgestellt. Untersuchungen zur Kopienzahl zeigten, daß im Makronukleus das HSP7O-Gen in einer Kopienzahl von 2000-2500 Kopien vorlag. Dies entsprach der durchschnittlichen Kopienzaht von E.crassus. Um Promotor-Elemente zu identifizieren, wurde der Transkriptionsstartpunkt bestimmt. Eine Analyse der 5'-nicht kodierten Region zeigte, daß keine Sequenzhomologie zu den bekannten core-Promotor-Elementen TATA-Box und Initiator-Element bestand. Durch Sequenzvergleiche mit der bekannten Konsensussequenz der Hitze-Schock-Elemente wurden jedoch zwei Hitze-Schock-Elemente identifiziert. Weitere Untersuchungen zur Expression des HSP7O-Gens nach Hitzeschock und ohne Hitzeschock erfolgten sowohl auf der Ebene der RNA als auch auf der Protein-Ebene. Durch eine Northern-Blot-Analyse wurde eine erhöhte Transkripmenge der HSP70-mRNA in hitzeinduzierten Ciliaten festgestellt. Auf der Protein-Ebene zeigte sich jedoch kein quantitativer Unterschied zwischen hitzeinduzierten und nicht hitzeinduzierten Zellen. Inhaltsverzeichnis: Inhaltsverzeichnis: 1.ZUSAMMENFASSUNG1 2.EINLEITUNG2 2.1Die Genomorganisation des hypotrichen Ciliat Euplotes crassus2 2.2Konjugation und Makronukleusentwicklung bei Euplotes crassus4 2.3Transk: riptinelle Regulation der Genexpression5 2.4Transkriptionelle Regulation der Genexpression am Beispiel der Hitzeschockgene7 2.5Ziel der Arbeit8 3.MATERIAL9 3.


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Product Details
  • ISBN-13: 9783838610085
  • Publisher: Diplom.de
  • Binding: Paperback
  • Language: German
  • Returnable: N
  • Spine Width: 5 mm
  • Width: 148 mm
  • ISBN-10: 3838610083
  • Publisher Date: 07 Aug 1998
  • Height: 210 mm
  • No of Pages: 88
  • Series Title: German
  • Weight: 127 gr


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