Expression and localization of Polo-like kinase 2 in osteosarcoma cells

Objective:To construct the expression plasmid of human Polo-like kinase 2(hPlk2) gene and identify the expression and localization of hPlk2 in osteosarcoma MG-63 cells.Methods: Human Plk2 coding sequence was obtained by polymerase chain reaction(PCR) amplification and cloned into 3* Flag vector.After the target region was identified by enzyme digestion and sequencing,the plasmid was transfected into osteosarcoma MG-63 cells.The expression of the recombinant plasmid in MG-63 cells was detected by Western blot.The localization of 3* Flag-hPlk2 in MG-63 cells was observed with laser scanning confocal microscopy.Purify the hPlk2 protein by immunoprecipitation assay.Results: hPlk2 was constructed into the expressing vector 3* Flag successfully.The length of the fragment identified by restriction enzyme digestion was 2058bp.The expression of 3* Flag-hPlk2 fusion protein with a molecular weight of 80kDa was detected by Western blot and pulled down by Flag antibody,and its localization was in the cytoplasm and perinucleus in MG-63 cells.Conclusion: The recombinant plasmid of hPlk2 gene was successfully cloned into eukaryotic expressing vector,and the expression of 3* Flag-hPlk2 fusion protein was identified and pulled down by Flag antibody,expressed in cytoplasm and perinucleus.