RPLPO gene encodes the acidic ribosomal phosphoprotein large P0 subunit, which is a component of the 60S subunit. The full-length cDNA sequence of porcine RPLPO was obtained from skeletal muscle of fetal pig cDNA library and deposited in GenBank. The nucleotide sequence and the predicted protein sequence shared high sequence identity with other mammalian homologues. A C/A single nucleotide substitution in exon 5 was detected as Csp6 Ⅰ polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) shows allele frequency diversity among Tongcheng, Xiaomeishan, Yushan, Large White, Landrace, and Duroc breeds. Analyses of somatic cell hybrid panel (SCI-IP) and radiation hybrid (IMpRH) panel showed that the RPLPO gene was mapped to SSC 14q22-q24 and was closely linked to locus SW1321 (25 cR, LOD = 14.54).