Gene cloning, expression and purification of fusion protein epidermal growth factor-linker-trichosanthin / 南方医科大学学报
Journal of Southern Medical University
;
(12): 205-207, 2007.
Artículo
en Chino
| WPRIM
| ID: wpr-298204
ABSTRACT
<p><b>OBJECTIVE</b>To construct a recombinant expression vector of the fusion protein epidermal growth factor (EGF)-Linker-trichosanthin (TCS) and achieve its expression in E. coli to obtain purified EGF-linker-TCS fusion protein.</p><p><b>METHODS</b>The gene fragments of EGF-linker were amplified by PCR and inserted into the expression plasmid PQE30-TCS, followed by transformation of the recombinant plasmid into E. coli M15 for expression of the fusion protein. Ni-FF column chromatography was utilized for purification of the expressed product.</p><p><b>RESULTS</b>The recombinant plasmid PQE30-EGF-linker-TCS was stably and highly expressed in E. coli M15. The expressed product existed in the form of soluble protein accounting for about 40% of total cellular protein and reached a purity of above 95% after purification with Ni-FF column chromatography.</p><p><b>CONCLUSION</b>The recombinant plasmid PQE30/EGF-linker-TCS has been successfully constructed, which provides a basis for further structural and functional study of EGF and TCS and their potential clinical application for cancer therapy.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes de Fusión
/
Tricosantina
/
Western Blotting
/
Clonación Molecular
/
Electroforesis en Gel de Poliacrilamida
/
Factor de Crecimiento Epidérmico
/
Escherichia coli
/
Vectores Genéticos
/
Genética
/
Metabolismo
Límite:
Humanos
Idioma:
Chino
Revista:
Journal of Southern Medical University
Año:
2007
Tipo del documento:
Artículo
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