Abstract: In present study, the luteinizing hormone betasubunit (LHβ) gene was selected as a candidate gene for the high prolificacy in Jining Grey goats. According to the sequence of ovine LHβ gene, six pairs of primers were designed to detect single nucleotide polymorphisms in the 5′ regulatory region, exon 1, exon 2 and exon 3 of ovine LHβ gene in high prolificacy breed (Jining Grey goat), medium prolificacy breed (Boer goat) and low prolificacy breeds (Liaoning Cashmere and Angora goats) by PCRSSCP method. The results showed that the similarity in nucleotide sequence of goat and ovine LHβ gene was 98%, and only the products amplified by primers P1 and P5 displayed polymorphisms. For primer P1, three genotypes (AA, AB and BB) were detected in all four goat breeds. Sequencing revealed that two nucleotide substitutions (202C→A and 210C→T) at the 5′regulatory region of LHβ gene in the BB genotype compared with the AA genotype. The differences of the litter size between different genotypes were nonsignificant in Jining Grey goats（P>0.05）. For primer P5, three genotypes (CC, CD and DD) were detected in Jining Grey goats, two genotypes (CC and CD) were detected in Liaoning Cashmere goats, and only one genotype CC was detected in Boer and Angora goats. Sequencing revealed one single nucleotide mutation (1124C→T) at exon 2 of LHβ gene in the DD genotype compared with the CC genotype. Genotype frequency of CC, CD and DD was 0.38, 0.44 and 0.18 in Jining Grey goats, respectively. The Jining Grey does with genotype DD or CD had 0.99 (P<0.01) or 0.87 (P<0.01) kids more than those with genotype CC. The fitness tests showed that the Jining Grey goat population was in HardyWeinberg equilibrium (P1 locus:χ2=1.66, P=0.437; P5 locus:χ2=1.22, P=0.544).