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Chinese Journal of Biotechnology ; (12): 35-39, 2002.
Artículo en Chino | WPRIM | ID: wpr-231378

RESUMEN

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 1:12,800, were indicated the specificity to prostate tissue.


Asunto(s)
Animales , Humanos , Ratones , Anticuerpos , Alergia e Inmunología , Formación de Anticuerpos , Antígenos de Superficie , Carboxipeptidasas , Genética , Alergia e Inmunología , Cromatografía de Afinidad , Métodos , Clonación Molecular , ADN Complementario , Genética , Escherichia coli , Genética , Expresión Génica , Vectores Genéticos , Glutamato Carboxipeptidasa II , Ratones Endogámicos BALB C , Estructura Terciaria de Proteína , Genética , Fisiología , Proteínas Recombinantes de Fusión , Genética , Alergia e Inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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