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.
Article
em Zh
| WPRIM
| ID: wpr-298204
Biblioteca responsável:
WPRO
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:
1
Base de dados:
WPRIM
Assunto principal:
Proteínas Recombinantes de Fusão
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Tricosantina
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Western Blotting
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Clonagem Molecular
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Eletroforese em Gel de Poliacrilamida
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Fator de Crescimento Epidérmico
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Escherichia coli
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Vetores Genéticos
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Genética
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Metabolismo
Limite:
Humans
Idioma:
Zh
Revista:
Journal of Southern Medical University
Ano de publicação:
2007
Tipo de documento:
Article