PCR-SSCP ANALYSIS OF GROWTH DIFFERENTIATION FACTOR-9 GENE OF GAROLE SHEEP

Growth differentiation factor-9 (GDF-9), one of the oocyte-derived members of the transforming growth factor - β super family has played an imperative role for follicular growth and ovulation. The present study was to investigate the polymorphism of exon-1 of GDF-9 gene in the most prolific Garole sheep breed. Blood samples were collected from 28 ewes and genomic DNA was extracted using the modified high salt method. The 462 bp amplified PCR product was analyzed for polymorphism by SSCP method. The results indicated two different banding patterns AA and AB for this fragment. The frequencies of AA and AB genotypes were 0.857 and 0.143. The allelic frequencies of A and B alleles were 0.928 and 0.072. The Calculated Chi-square value for exon-1 of GDF-9 gene (0.032) was found to be lesser than that of tabulated values at 5% and 1% level of significance indicating that the population under study was in Hardy-Weinberg Equilibrium. The 462 bp nucleotide sequence was subjected to BLAST analysis. Phylogenetic analysis revealed that nucleotide sequence of studied sample of Garole sheep is 100 percent similar with the published sequence of sheep and form a common cluster indicating their evolutionary closeness.