The coding region of PKM2 was cloned by RT-PCR, and was subjected to sequence bioinformatics analysis, tissue expression profiling and subcellular localization. The results showed that the CDS of porcine PKM2 gene was 1 596 bp in length, which encodes 531 amino acids with the molecular weight 5.79×104 , the isoelectric point of 7.96, and multiple phosphorylation sites. PKM2 protein had two domains, pyruvate kinase, barrel domain and pyruvate kinase, alpha/beta domain. Regarding to secondary structure, the α-helices and random coil accounted for the main types in the PKM2 protein. The PKM2 protein sequence was very conserved among species, and phylogenetic tree analysis showed that the procine PKM2 protein is closely related to rabbits. The tissue expression profiling revealed that PKM2 gene was highly expressed in kidney and small intestine, and was relatively low expressed in liver and muscle. Subcellular localization showed that the PKM2 protein is expressed in the cytoplasm in porcine 3D4/21 and PK15 cells but not in the nucleus.