Leishmania cell-free protein expression system.
Methods
; 55(1): 58-64, 2011 Sep.
Article
em En
| MEDLINE
| ID: mdl-21704167
ABSTRACT
Cell-free protein expression is an important tool for a rapid production, engineering and labeling of recombinant proteins. However the complex protocols for preparation of eukaryotic cell-free protein expression systems result in high manufacturing costs and limit their utility. Recently we reported a novel cell-free expression system based on the lysate of a fermentable protozoan Leishmania tarentolae. Herein we describe a protocol for high throughput protein expression using Leishmania cell-free lysate. The protocol combines PCR-based synthesis and engineering of translation templates with a combined transcription-translation system. The protocol is adapted to multiwell plate format and allows translation of large protein libraries. In the presented example we translate in vitro and isolate a nearly complete complement of mammalian Rab GTPases. Further applications and developments of the system are discussed.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes de Fusão
/
Engenharia de Proteínas
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Sistema Livre de Células
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Reação em Cadeia da Polimerase
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Proteínas rab de Ligação ao GTP
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Ensaios de Triagem em Larga Escala
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Leishmania
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Methods
Ano de publicação:
2011
Tipo de documento:
Article