Detection of Polymorphism in Exon 2 of GDF9 gene in Pure and Crossbred of Pakistani Sheep

GDF9 gene is one of the most important effective factors on litter size in sheep. Thus, the aim of the present study was to detect single nucleotide polymorphisms (SNPs) available in exon 2 of GDF9 gene in pure and crossbred of Pakistani sheep using PCR-SSCP. Hence, blood samples were collected from 30 Pakistani sheep, 17 crossbred sheep (Pakistani rams × LoriBakhtiari) and 7 crossbred sheep (Pakistani rams × Kermani ewes) and genomic DNA was extracted using salting-out method. A 634 bp fragment was amplified using one specific primer pairs using polymerase chain reaction. After single stranded conformation polymorphism (SSCP) of PCR products, band patterns of GDF9 gene were observed on 8% polyacrylamide gel stained with silver-nitrate method. In total, 4 banding patterns 1, 2, 3 and 4 were obtained with frequencies of 0.193, 0.129, 0.371 & 0.407, respectively. The sequencing results were led to identification of 3 mutations in 443, 477 and 721 nucleotide situations of GDF9 gene in the studied populations.