Study on the cloning, expression and identification of the renal cell carcinoma-associated antigen G250/MN/CA Ⅸ / 中华泌尿外科杂志

ABSTRACT

Objective To report the cloning, expression and identification of the tumor-associated antigen G250/MN/CA Ⅸ. Methods The total RNA was extracted from renal cell carcinoma tissue samples from 54 male patients. Gene fragments encoding G250 was obtained by RT-PCR,and was cloned into prokaryotic expression vector pET22b(+) and expressed in E.coli BL21(DE3);and the results were examined by SDS-PAGE gel electrophoresis. The recombinant protein was studied by Western blot test. Results DNA sequence analysis showed that the obtained sequence was the same as that showed in GenBank.Gene of G250 was expressed in E.coli BL21 successfully. Western blot analysis showed that the recombinant protein could be specially recognized by monoclonal antibody. It had better antigenicity and specificity. Conclusions This study provides experimental basis for the purification of G250/MN/CA Ⅸ protein and the further study of G250/MN/CA Ⅸ function and preparation of antibody.