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.
Article
em Zh
| WPRIM
| ID: wpr-231378
Biblioteca responsável:
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 1:12,800, were indicated the specificity to prostate tissue.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Fisiologia
/
Proteínas Recombinantes de Fusão
/
Carboxipeptidases
/
Expressão Gênica
/
Cromatografia de Afinidade
/
Clonagem Molecular
/
Estrutura Terciária de Proteína
/
DNA Complementar
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Glutamato Carboxipeptidase II
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2002
Tipo de documento:
Article