Expression of human tumstatin in Pichia pastoris and its bioactivity / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 451-456, 2006.
Artículo
en Chino
| WPRIM
| ID: wpr-286268
ABSTRACT
Human tumstatin(hTumstatin)cDNA was amplified from recombinant plasmid pET-3c-tum, cloned in frame with the signal sequence in yeast vector pPICZalphaA and transformed into Pichia pastoris GS115 by electroporation. The expression of hTumstatin in GS115(pPICZalpha-tum)was then induced by methanol and secreted into the culture medium, with a yield of 25mg/L as shown by SDS-PAGE and Western blotting. The expressed hTumstatin was purified to more than 85% purity using a simple one-step SP-Sepharose cation exchange chromatography. The MTT and chick chorioallantoic membrane assay showed that the yeast produced hTumstatin could inhibit the proliferation of human umbilical vein endothelial cells and the neovascularization induced by bFGF. Hoechst 33258 fluorescent staining also demonstrated the apoptotic change in endothelial cellular nuclear morphology.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Pichia
/
Autoantígenos
/
Cordón Umbilical
/
Proteínas Recombinantes
/
Células Cultivadas
/
Electroporación
/
ADN Complementario
/
Inhibidores de la Angiogénesis
/
Biología Celular
/
Colágeno Tipo IV
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2006
Tipo del documento:
Artículo
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