Reporter Vector Construction of Human ODF Gene Promoter

Objective: To construct a specific reporter vector to monitor the activity of human osteoclast differentiation factor (ODF) gene promoter. Methods: Two DNA segments of ODF gene promoter (-2 433 - -1 243 bp and -1 262 - +100 bp) were amplified by PCR from human genome DNA and cloned into clone vector pGEM-T Easy. The gene fragments were correctly connected to the promoter without vector pEGFP-1 by restriction enzyme. Recombinant plasmid pEGFP-1-ODF was transferred into osteoblastic cell line UMR106. The expression of EGFP was detected under the monitor of human ODF gene promoter (-2 358-+100 bp). Results: pEGFP-1-ODF was the same as the design confirmed by restriction digestion and sequence analysis. The expressing EGFP could be detected in UMR106 by pEGFP-1-ODF transfection. Conclusion: Human ODF gene promoter reporter vector is significant for further study on ODF gene expression transfection level.