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
in Chinese
| WPRIM
| ID: wpr-231378
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Physiology
/
Recombinant Fusion Proteins
/
Carboxypeptidases
/
Gene Expression
/
Chromatography, Affinity
/
Cloning, Molecular
/
Protein Structure, Tertiary
/
DNA, Complementary
/
Reverse Transcriptase Polymerase Chain Reaction
/
Glutamate Carboxypeptidase II
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
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