Aim To construct recombinant plasmid pET32a-apolipoprotein AV and purify the protein with His tag. Methods The total RNA was extracted from tree shrew liver tissues, cDNA was then obtained by reverse transcription-polymerase chain reaction. Apolipoprotein AV gene fragment was amplified by PCR. The amplified products and pET32a plasmid were digested by restriction enzymes Xho I and Eco RI, and then the purified products were ligated by T4 DNA ligase. The recombinants were transformed into E.coli Top10 and BL21（DE3）. The apolipoprotein AV was induced with isopropy-β-D-thiogalactoside. The expressed conditions were optimized and purified by Nickel ion chelating resin. Purity analysis of the apolipoprotein AV was obtained by SDS-PAGE. Results Recombinant plasmid pET32a-apolipoprotein AV was successfully constructed and expressed in BL21（DE3）. Isopropy-β-D-thiogalactoside-induced target protein （about 60 kDa） was detected. The purity of recombinant tree shrew apolipoprotein AV was greater than 95%.