Your browser doesn't support javascript.
loading
Prokaryotic expression and identification of dual-fluorescence fusion proteins of small ubiquitin-like modifier and sentrin-specific protease / 生物工程学报
Chinese Journal of Biotechnology ; (12): 701-707, 2009.
Artículo en Chino | WPRIM | ID: wpr-286654
ABSTRACT
We cloned genes of four sentrin-specific protease (SENP), three small ubiquitin-like modifiers (SUMO), enhanced cyan fluorescent protein (ECFP) and yellow fluorescent protein (EYFP) by two-step PCR. Then we constructed expression vector B28 for SENP and B13 for ECFP-SUMO-EYFP. The recombinant plasmids were transformed into Escherichia coli BL21 and expression was induced by isopropyl beta-D-thiogalactoside, then recombinant proteins were purified by Ni-NTA agarose column ion-exchange chromatography. The proteins were analyzed with SDS-PAGE and identified with Western blotting. Except that SENP3 catalytic domain (SENP3C) truncated in the C termini and SENP5C expressed in inclusion body, others were expressed as soluble proteins. SDS-PAGE analysis showed that the relative molecular mass of these fusion proteins were consistent with theoretical ones, and the specificity of the fusion proteins were confirmed with Western blotting. The fusion proteins of SENP and ECFP-SUMO-EYFP can be successfully expressed in prokaryotic expression system. It lays the foundation for the fluorescence resonance energy transfer analysis.
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Endopeptidasas / Proteínas Bacterianas / Proteínas Recombinantes de Fusión / Cisteína Endopeptidasas / Dominio Catalítico / Proteína SUMO-1 / Proteínas Fluorescentes Verdes / Escherichia coli / Vectores Genéticos / Genética Límite: Humanos Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2009 Tipo del documento: Artículo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Endopeptidasas / Proteínas Bacterianas / Proteínas Recombinantes de Fusión / Cisteína Endopeptidasas / Dominio Catalítico / Proteína SUMO-1 / Proteínas Fluorescentes Verdes / Escherichia coli / Vectores Genéticos / Genética Límite: Humanos Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2009 Tipo del documento: Artículo