Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 1 de 1
Filter
1.
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 1:12,800, were indicated the specificity to prostate tissue.


Subject(s)
Animals , Humans , Mice , Antibodies , Allergy and Immunology , Antibody Formation , Antigens, Surface , Carboxypeptidases , Genetics , Allergy and Immunology , Chromatography, Affinity , Methods , Cloning, Molecular , DNA, Complementary , Genetics , Escherichia coli , Genetics , Gene Expression , Genetic Vectors , Glutamate Carboxypeptidase II , Mice, Inbred BALB C , Protein Structure, Tertiary , Genetics , Physiology , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Reverse Transcriptase Polymerase Chain Reaction
SELECTION OF CITATIONS
SEARCH DETAIL