Desenvolvimento de uma construção de gene repórter para a localização subcelular

Name: Desenvolvimento de uma construção de gene repórter para a localização subcelular
Brand: Edicoes Nosso Conhecimento
SKU: 9786630022315
Price: 7340.0 INR
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ISBN13: 9786630022315
Binding: Paperback
Subject: N/A
Publisher: Edicoes Nosso Conhecimento
Publisher Imprint: Edicoes Nosso Conhecimento
Publication Date: May 31, 2026
Pages: 88
Original Price: GBP 58.0
Language: Portuguese
Edition: N/A
Item Weight: 127 grams
BISAC Subject(s): Life Sciences / Genetics & Genomics

About The Book

A RNA pirofosfohidrolase (RppH) inicia a degrada��o das transcri��es atrav�s da remo��o do pirofosfato da extremidade 5' dos mRNAs, o que permite a liga��o da RNase E/RNase J nas bact�rias. Uma suposta RNA pirofosforilase est� presente nas c�lulas de C. reinhardtii e v�rias evid�ncias sugerem que a prote�na est� envolvida na degrada��o do mRNA no cloroplasto. A fim de determinar a localiza��o da prote�na nas c�lulas de Chlamydomonas, a extremidade 5' do gene rppH foi marcada com uma prote�na fluorescente verde com cod�es otimizados e introduzida nas c�lulas de Chlamydomonas. A GFP (Zsgreen 1) foi otimizada em termos de c�dons utilizando uma base de dados de utiliza��o de c�dons. O quadro de leitura da GFP sint�tica otimizada foi ajustado e clonado no vetor pBluescript-5'rppH SK+. O fragmento do gene 5'rppH-GFP foi ent�o clonado em tr�s vetores. Todas as constru��es foram verificadas. Os transformantes foram selecionados quanto � presen�a do gene GFP quim�rico por PCR. As experi�ncias de triagem levaram-nos a escolher um transformante que pudesse ser utilizado para trabalhos futuros. Al�m disso, embora a fluoresc�ncia n�o tenha sido verificada, a GFP sint�tica otimizada foi expressa em c�lulas de E. coli (BL21DE3), confirmando a funcionalidade da constru��o da GFP sint�tica in vivo.

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