Currently, the primary thrust of research in animal genetics is the identification of gene (so-called major genes), which after the expression of quantitative traits markedly. One of the major genes is CYP19 gene which is related to reproduction and production traits in livestock. This study aimed to evaluate the genetic polymorphism within CYP19 gene in Sahiwal cattle using PCR-RFLP technique. Genomic DNA extracted from 60 healthy cows was amplified using primer that were designed from the cattle CYP19 gene sequences. The polymorphism in CYP19 gene was detected using PCR-RFLP method and PvuII restriction enzyme. The frequency of genotype AA was found to be 48.33%, AB was 41.66% and BB was 10.0% respectively. The allele A allele was more frequent (0.69) than the allele B (0.31). In population was found deviations from Hardy-Weinberg Equilibrium (HWE).