Abstract

Different mutations in the bone morphogenetic protein 15 (BMP15) and the Growth Differentiation Factor 9 (GDF9) genes cause increased ovulation rate and infertility in a dosage-sensitive manner in sheep. In this study, blood samples (140 ewes and 10 rams) were initially taken from 150 Sangsari sheep breed in Damghan animal breeding Centre using venojects treated with the anti-clot substance (EDTA) and subsequently their DNA content were salted out and extracted. Using two pairs of specific primers, two DNA fragments were amplified from exon 1 of GDF-9 (462 bp) and exon 2 of BMP15 (141 bp) genes. The resulted PCR products were digested using HhaІ and HinfI restriction enzymes for GDF9 and BMP15 genes, respectively. Digested PCR products with HhaІ enzyme showed a G to A substiuation in GDF9 locus. The wild type allele of this gene (G/+) with two restriction site resulted DNA fragments of 156.52 and 254 bp while the mutant allele (G/-) with one restriction site resulted two DNA fragments with the size of 52 and 410 bp. Genotype frequencies for G (+/+),G (+/-) and G (-/-) were 70.72, 36.88 and 1.40%, respectively. Restriction digested of PCR products for BMP15 locus with Hinf I enzyme showed C to T transition. BMP15 luci was not polymorphic. From studied luci, only GDF9 was polymorphic in Iranian Sanghsari sheep.

Key words: PCR, polymorphism, GDF9, BMP15, Sangsari sheep.