Prolactin (PRL) regulates various functions in pigs including reproduction, mammary development and lactation. We used 5'-rapid amplification of cDNA ends (5'-RACE) to clone three full-length alleles of the porcine PRL receptor (pPRLR) from Landrace (alleles LR2 and LR4) and Yucatan miniature (MP) pigs, corresponding to the A and B alleles previously reported to be associated with reproductive traits. When expressed in Chinese hamster ovary (CHO-K1) cells, all three pPRLRs transduced differentiation signals to a beta-casein promoter with the same effectiveness, where human growth hormone (hGH) and porcine PRL (pPRL) were more effective ligands than ovine PRL (oPRL). The pPRLR had a lower binding affinity for oPRL than pPRL while binding affinity for hGH was not different between the three pPRLR variants. The pPRLRs primarily localized to the cytoplasm with perinuclear concentration. In conclusion, we have cloned three allelic variants of the pPRLR and have functionally characterized these as different from the hPRLR. However, our data do not support the proposal that allelic variation of the pPRLR confers functional differences in vivo.