Construction of eukaryotic expression vector of human miR-34a and it's expression

OBJECTIVE:To construct a recombinant eukaryotic expression plasmid of miR-34a and express it in osteosarcoma SOSP-9607 cells. METHODS:miR-34a precursor sequence was amplified by PCR from the human genomic DNA,and then cloned into the pcDNA 3.1(+) eukaryotic expression vector to produce the recombinant pcDNA-miR34a. The osteosarcoma SOSP-9607 cells were transfected with recombinant pcDNA-miR34a,and then selected to obtain the stable cells,and then the miR-34a expression was verified by Northern blot and real time RT-PCR. RESULTS:The miR-34a eukaryotic expression vector was successfully constructed. pcDNA-miR34a could up-regulate the expression of miR-34a in osteosarcoma SOSP-9607 cells. The miR-34a high expression osteosarcoma cells were obtained. CONCLUSION:The eukaryotic expression plasmid of miR-34a is successfully constructed and effectively expressed in osteosarcoma SOSP-9607 cells,which facilitats the further study of miR-34a fuction in osteosarcoma.