Detection polymorphism in GDF9, BMP15 and FecB genes in Sangsari sheep breed of Iran the using PCR-RFLP and PCR-SSCP

Detection polymorphism in GDF9, BMP15 and FecB genes in Sangsari sheep breed of Iran the using PCR-RFLP and PCR-SSCP

Phenotypic evaluation and making culling decision of candidate animals for the traits under consideration through applying classic genetics are usually a costly task and require much time to be carried out. Selection based upon markers could result in an increased accuracy as well as selection response of animals. In this study, blood samples (140 ewes and 20 rams) were initially taken from 160 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 three pairs of specific primers, three DNA fragments were amplified from exon 1 of GDF-9 (462bp), exon 2 of BMP15 (141bp) and FecB (190bp) genes. After the extraction and undertaking quantitative and qualitative tests (including spectrophotometer and gel agarose 8%) the required amount of DNA for polymerase chain reaction (PCR) were determined The resulted PCR products were digested using HhaІ, HinfI and AvaП restriction enzymes for GDF9, BMP15 and FecB genes, respectively Digested PCR products with HhaІ enzyme showed a G to A substitution in GDF9 locus. The wild type allele of this gene (G/+) with two restriction site resulted DNA fragments of 156, 52 and 254bp while the mutant allele (G/-) with one restriction site resulted two DNA fragments with the size of 52 and 410bp. 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 and FecB luci were not polymorphic. From studied luci, only GDF9 was polymorphic in Iranian Sangsari sheep.

Phenotypic evaluation and making culling decision of candidate animals for the traits under consideration through applying classic genetics are usually a costly task and require much time to be carried out. Selection based upon markers could result in an increased accuracy as well as selection response of animals. In this study, blood samples (140 ewes and 20 rams) were initially taken from 160 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 three pairs of specific primers, three DNA fragments were amplified from exon 1 of GDF-9 (462bp), exon 2 of BMP15 (141bp) and FecB (190bp) genes. After the extraction and undertaking quantitative and qualitative tests (including spectrophotometer and gel agarose 8%) the required amount of DNA for polymerase chain reaction (PCR) were determined The resulted PCR products were digested using HhaІ, HinfI and AvaП restriction enzymes for GDF9, BMP15 and FecB genes, respectively Digested PCR products with HhaІ enzyme showed a G to A substitution in GDF9 locus. The wild type allele of this gene (G/+) with two restriction site resulted DNA fragments of 156, 52 and 254bp while the mutant allele (G/-) with one restriction site resulted two DNA fragments with the size of 52 and 410bp. 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 and FecB luci were not polymorphic. From studied luci, only GDF9 was polymorphic in Iranian Sangsari sheep.