[Expression and purification of the N-domain of human canstatin and its bioactivity].
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
; 35(5): 483-7, 2003 May.
Article
en Zh
| MEDLINE
| ID: mdl-12766812
ABSTRACT
Total RNA was extracted from placenta umbilical tissue. The canstatin cDNA was amplified from total RNA by net-RT-PCR technique and cloned into pSP72, and the resulted plasmid pSP72C was used as template to amplify its N-domain. The amplified 1-89 aa N-domain was then cloned into pET-3c. The resulted plasmid pET-CN was transformed into E. coli BL21(DE3). The N-domain was efficiently expressed after IPTG induction as a 10 kD band on SDS-PAGE. The expressed product accounted for approximately 35.3% of the total bacterial proteins, as estimated by densitometry and existed mainly as inclusion body. The inclusion bodies were washed, lysed and the reactivated proteins were purified by the Sephadex G-100 gel filtration to a purity of 92.6%. CAM assay showed that N-domain effectively inhibited the angiogenesis of chicken embryo microcapillary vessel.
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Base de datos:
MEDLINE
Asunto principal:
Placenta
/
Proteínas Gestacionales
/
Inhibidores de la Angiogénesis
Límite:
Humans
Idioma:
Zh
Revista:
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
Año:
2003
Tipo del documento:
Article
País de afiliación:
China