Abstract: The aim of this study was to construct a porcine pLV-miR-183 lentiviral vector and detect its expression in porcine kidney cells（PK15）. The genomic DNA of Guangxi Bama mini-pig was used as the template to clone the precursor and part of flanking sequences of miR-183 using PCR, and the sequence was inserted into pLV-CMV-MCS-EF1 vector by T4 ligase to construct the pLV-miR-183 lentiviral expression vector. The positive recombinant plasmids of miR-183 identified by PCR and sequencing were then transfected into PK15 cells, and the transfection efficiency was measured by fluorescence inverted microscope. The results showed that the length of miR-183 amplification sequence was 329 bp, which was consistent with the reference sequence. After transfection of positive recombinant plasmid pLV-miR-183 into PK15 cells for 48 h, strong expression of green fluorescent protein was detected under fluorescence microscopy, and its high expression level was verified by qRT-PCR,indicating that the constructed pLV-miR-183 lentiviral vector had high expression activity in PK15 cells. This study successfully constructed the porcine miR-183 lentiviral expression vector, which laid a certain theoretical foundation for the further study on the mechanism of disease action of miR-183.