Entwicklung eines Reportergen-Konstrukts zur Bestimmung der subzellulären Lokalisierung

Name: Entwicklung eines Reportergen-Konstrukts zur Bestimmung der subzellulären Lokalisierung
Brand: Verlag Unser Wissen
SKU: 9786630009514
Price: 7340.0 INR
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by Biruk Abrha

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ISBN13: 9786630009514
Binding: Paperback
Subject: N/A
Publisher: Verlag Unser Wissen
Publisher Imprint: Verlag Unser Wissen
Publication Date: May 31, 2026
Pages: 92
Original Price: GBP 58.0
Language: German
Edition: N/A
Item Weight: 137 grams
BISAC Subject(s): Life Sciences / Genetics & Genomics

About The Book

Die RNA-Pyrophosphohydrolase (RppH) leitet den Abbau von Transkripten ein, indem sie Pyrophosphat vom 5'-Ende der mRNAs entfernt, was die Bindung von RNase E/RNase J in Bakterien erm�glicht. In Zellen von C. reinhardtii ist eine mutma liche RNA-Pyrophosphorylase vorhanden, und mehrere Hinweise deuten darauf hin, dass das Protein am mRNA-Abbau im Chloroplasten beteiligt ist. Um den Ort des Proteins in Chlamydomonas-Zellen zu bestimmen, wurde das 5'-Ende des rppH-Gens an ein codonoptimiertes gr�n fluoreszierendes Protein (GFP) gekoppelt und in Chlamydomonas-Zellen eingef�hrt. GFP (Zsgreen 1) wurde unter Verwendung einer Codon-Usage-Datenbank codonoptimiert. Der Leserahmen des optimierten synthetischen GFP wurde angepasst und in den Vektor pBluescript-5'rppH SK+ kloniert. Das 5'rppH-GFP-Genfragment wurde anschlie end in drei Vektoren kloniert. Alle Konstrukte wurden verifiziert. Die Transformanten wurden mittels PCR auf das Vorhandensein des chim�ren GFP-Gens untersucht. Die Screening-Experimente f�hrten dazu, dass wir einen Transformanten ausw�hlten, der f�r die weitere Arbeit verwendet werden konnte. Obwohl die Fluoreszenz nicht �berpr�ft wurde, wurde das optimierte synthetische GFP in E. coli (BL21DE3)-Zellen exprimiert, was die Funktionalit�t des synthetischen GFP-Konstrukts in vivo best�tigte.

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Entwicklung eines Reportergen-Konstrukts zur Bestimmung der subzellulären Lokalisierung

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