Sviluppo di un costrutto genico reporter per la localizzazione subcellulare

Name: Sviluppo di un costrutto genico reporter per la localizzazione subcellulare
Brand: Edizioni Sapienza
SKU: 9786630017199
Price: 7340.0 INR
Availability: InStock

by Biruk Abrha

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ISBN13: 9786630017199
Binding: Paperback
Subject: N/A
Publisher: Edizioni Sapienza
Publisher Imprint: Edizioni Sapienza
Publication Date: May 31, 2026
Pages: 88
Original Price: GBP 58.0
Language: Italian
Edition: N/A
Item Weight: 127 grams
BISAC Subject(s): Life Sciences / Genetics & Genomics

About The Book

L'RNA pirofosfidrolasi (RppH) avvia la degradazione dei trascritti rimuovendo il pirofosfato dall'estremit� 5' degli mRNA, consentendo cos� il legame con RNasi E/RNasi J nei batteri. Una putativa RNA pirofosforilasi � presente nelle cellule di C. reinhardtii e diverse evidenze suggeriscono che la proteina sia coinvolta nella degradazione dell'mRNA nel cloroplasto. Al fine di determinare la localizzazione della proteina nelle cellule di Chlamydomonas, l'estremit� 5' del gene rppH � stata marcata con una proteina fluorescente verde ottimizzata a livello di codoni e introdotta nelle cellule di Chlamydomonas. La GFP (Zsgreen 1) � stata ottimizzata a livello di codoni utilizzando un database sull'utilizzo dei codoni. Il frame di lettura della GFP sintetica ottimizzata � stato regolato e clonato nel vettore pBluescript-5'rppH SK+. Il frammento genico 5'rppH-GFP � stato quindi clonato in tre vettori. Tutti i costrutti sono stati verificati. I trasformanti sono stati sottoposti a screening per la presenza del gene GFP chimerico mediante PCR. Gli esperimenti di screening ci hanno portato a scegliere un trasformante che potesse essere utilizzato per ulteriori lavori. Inoltre, sebbene la fluorescenza non sia stata verificata, la GFP sintetica ottimizzata � stata espressa in cellule di E. coli (BL21DE3), confermando la funzionalit� del costrutto GFP sintetica in vivo.

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