The recombinant expression plasmid pGEX hSDCT2 was constructed and introduced into E.coli. GST hSDCT2 fusion protein was expressed and purified by GST Sepharose 4B affinity chromatography. The purified fusion protein was used as an immunogen to inoculate rabbits. Then the multi clonal antibodies against GST hSDCT2 fusion protein were obtained. Northern blotting showed two transcripts (3 6 kb and 4 3 kb) of hSDCT2 gene in different human tissues. The 3 6 kb hSDCT2 gene transcript was almost not expressed in heart, skeletal muscle, thymus, small intestine, lung and peripheral blood leukocyte, and slightly expressed in brain, colon, spleen, liver and placenta, but highly expressed in kidney. In addition, the 4 3 kb transcript was detected in kidney and spleen. Western blotting confirmed that hSDCT2 protein was expressed in the tissues mentioned above in similar manner. Immunohistochemical double staining demonstrated that hSDCT2 was mainly expressed on the basolateral membrane of renal proximal tubular, which was different from hSDCT1 expressed on the brush border membrane of renal proximal tubular. These results provide a foundation for further research on structure and function of hSDCT2 gene.
Key words
human sodium dependent dicarboxylate transporter2 /
antibody /
tissue expression profile
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