The present study was performed to investigate the polymorphism in the ATP-binding cassette superfamily G member 2 transporter (ABCG2) gene in Sahiwal cattle by PCR-RFLP assay. PCR-RFLP analysis of exon 14 was carried out using restriction enzyme PstI. PstI-RFLP for exon 14 revealed monomorphic pattern showing only one type of genotype, AA (292 bp) with genotypic frequencies of 1.0. AC and CC genotype were not observed in the studied Sahiwal population. The present study revealed that the selected Sahiwal population is monomorphic for ABCG2 gene as only single genotype (AA) was found in all the animals which were screened for studying polymorphism in ABCG2 gene.