Construction of pHsa-m122 Expression Vector and Determination of Its Expression Activity

Objective To construct and analyze the activity of the expression vector pHsa-m122. Method Human precursor microRNA (miR-122) was obtained from the genomic DNA by PCR. Restriction sites and poly-T were introduced by ligation. The fragment was then inserted into the eukaryotic inducible expression vector pSuper. The insert was verified by restriction endonuclease digestion and sequencing, and the gene expression was evaluated by fluorescence microscopy, flow cytometry and Western Blot. Result miR-122 was successfully cloned into the eukaryotic expression vector pSuper. The expression product is biologically active. Conclusion The eukaryotic expression vector pHsa-m122 was successfully constructed. The expression vector may be used in the future study of hepatitis virus replication and the development of liver cancer.