Cloning and expression of extracellular domain of prostate specific membrane antigen in Escherichia coli and preparation of polyclonal antibody / 生物工程学报
Chinese Journal of Biotechnology
; (12): 35-39, 2002.
Artículo
en Chino
| WPRIM (Pacífico Occidental)
| ID: wpr-231378
Biblioteca responsable:
WPRO
ABSTRACT
Human Prostate Specific Membrane Antigen(PSMA) cDNA was amplified using total RNA extracted from prostate carcinoma tissue by RT-PCR. The cDNA fragment of extracellular domain of PSMA(edPSMA) gene was amplified by PCR and cloned into expression vector pMAL-c2x. Sequence analysis of both PSMA and edPSMA revealed identity to the GenBank reported. The edPSMA was expressed in E. coli as part of a fusion protein with MBP as the induction of IPTG. Western blot analysis showed the recombinant protein could react with PSMA monocloned antibodies 4G5. MBP-edPSMA fusion protein were purified by amylose resin affinity chromatography and showed to be homogeneity in SDS-PAGE(120 kD). BALB/C mice were immunized with the purified protein for the preparation of polyclonal antibody. The polyclonal antibody, which had a title of 112,800, were indicated the specificity to prostate tissue.
Texto completo:
Disponible
Contexto en salud:
Enfermedades Desatendidas
Problema de salud:
Enfermedades Desatendidas
/
Zoonosis
Base de datos:
WPRIM (Pacífico Occidental)
Asunto principal:
Fisiología
/
Proteínas Recombinantes de Fusión
/
Carboxipeptidasas
/
Expresión Génica
/
Cromatografía de Afinidad
/
Clonación Molecular
/
Estructura Terciaria de Proteína
/
ADN Complementario
/
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Glutamato Carboxipeptidasa II
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2002
Tipo del documento:
Artículo