One-step purification of chimeric green fluorescent protein providing metal-binding avidity and protease recognition sequence.
Asian Pac J Allergy Immunol
;
2003 Dec; 21(4): 259-67
Artigo
em Inglês
| IMSEAR
| ID: sea-37082
ABSTRACT
Gene fusion technique was successfully applied as a potential approach to create a metal-binding site to assist one-step purification of green fluorescent protein (GFP). The chimeric GFP carrying hexapolyhistidine (H6GFPuv) was purified to homogeneous protein via the Immobilized Metal Affinity Chromatography charged with zinc ions. Removal of metal tagger could readily be performed by using enterokinase enzyme. Engineering of the hexahistidine and enterokinase cleavage sites (DDDDK) onto the chimeric protein did not significantly affect the fluorescent property and the binding avidity to Burkholderia pseudomallei protease of a chimeric protease-binding GFP (H6PBGFPuv). This concludes that engineering of repetitive histidine regions onto interested target protein along with the enterokinase cleavage sites will ease the complication of protein purification.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Peptídeo Hidrolases
/
Zinco
/
Sítios de Ligação
/
Proteínas Recombinantes de Fusão
/
Enteropeptidase
/
Engenharia Genética
/
Cromatografia de Afinidade
/
Burkholderia pseudomallei
/
Proteínas de Fluorescência Verde
/
Eletroforese em Gel de Poliacrilamida
Tipo de estudo:
Estudos de avaliação
Idioma:
Inglês
Revista:
Asian Pac J Allergy Immunol
Ano de publicação:
2003
Tipo de documento:
Artigo
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