Kisspeptins receptor (KISS1R), also called GPR54, is a key regulator of puberty in many species. KISS1R and its genetics in pigs remain unexplored. The objective of this study was to characterize the porcine KISS1R gene and evaluate the association of KISS1R mutations with age at puberty in sows. KISS1R was assigned to pig chromosome 2q21-24 by radiation hybrid mapping. It has a 1438 bp full-length cDNA and spans 3349 bp genomic sequence consisting of five exons and four introns. Semi-quantitative RT-PCR showed that KISS1R transcripts was particularly abundant in the adrenal, prostate, testis, thymus, pituary and hypothalamus. KISS1R mRNA content in the hypothalamus was determined by real-time quantitative RT-PCR, and it fluctuated during the oestrous cycle with the highest level in the luteal phase. Anoestrus sows had markedly lower hypothalamic KISS1R mRNA content than cyclic animals. Seven KISS1R SNPs were identified in the founder animals of a White Duroc x Erhualian intercross. One missense mutation (T/C(245)) showed quite different allele distribution in Chinese and Western breeds. All F(0), F(1) animals and 367 detailed phenotyped cyclic F(2) sows in the White Duroc x Erhualian intercross were genotyped for three KISS1R polymorphisms. No significant association of KISS1R haplotypes and haplotype pairs with age at puberty was observed in the resource population, indicating that mutations in KISS1R are not responsible for divergent age at puberty in White Duroc and Erhualian pigs.