Abstract: The mRNA differential display technique was performed to investigate gene expression differences in the longissimus dorsi muscle from Wujin and Large white pigs. A fragment of one differentially expressed gene was isolated and sequenced. A complete cDNA sequence of the gene was obtained using the Rapid Amplification of Cdna Ends (RACE) method. The open reading frame of this gene encodes a protein of 428 amino acids which is homologous with the 6-Phosphofructo-2-Kinase/Fructose-2,6-Biphosphatase 1 (PFKFB1) of 6 species: cattle (97%), human (97%), horse (96%), rhesus monkey (96%), rat (94%) and mouse (94%). This newly identified gene was respectively defined as the swine PFKFB1 gene and had been assigned GeneID: 100233197. The phylogenetic tree analysis revealed that the swine PFKFB1 gene has a closer genetic relationship with the PFKFB1 gene of cattle. The tissue expression analysis indicated that the swine PFKFB1 gene has a broad tissue distribution. The experiment is the first to establish the primary foundation for further research on the swine PFKFB1 gene.