Selenoprotein genes contain selenium in the form of selenocysteine which is involved in protecting the cells from oxidative stress. Soils in India differ greatly in selenium concentrations affecting feed stuffs for selenium availability. Selenoproteins have recently been identified in variety of living organisms including humans which have 25 selenoprotein genes. Among these families of selenoprotein genes, we sequenced Gpx1 gene (Glutathione peroxidases1) and Dio1 gene (Iodothyronine deiodinases) in Bubalus bubalis. Gpx1 is most abundant and ubiquitously expressed selenoprotein which helps to protect against the damaging effects of hydrogen peroxide and oxygen rich free radicals whereas Dio1 is expressed mainly in liver, thyroid gland and adipose tissue, its main function is to convert tetraiodothyronine (T4) to its active form thyroxine (T3) in the presence of deiodinases enzyme. The main aim of the study was to characterize these two genes and to find out the buffalo specific SNPs. This was accomplished by designing primers using cattle database and sequencing a panel of 24 samples consisting of 6 diverse breeds of buffalo. Gpx1 consisted of 2 exons (Accession ID: JQ031269) while Dio1 comprised 4 exons (Accession ID: JQ791197). In Gpx1 gene, 9 SNPs were recorded and 4 were non synonymous, changing amino acid were distributed equally in both exon. In exon 1, A141G (aa Q5R) and G161A (aa A12T); and in exon 2 C785T (aa R132W) and A808T (aa S139R). In Dio1 gene, 3 non synonymous SNPs were identified at A188G (aa H22R), C215G (aa T31R) and G941A (aa V146I). These SNPs are novel and reported for the first time in Indian buffalo and has a potential for their use in diversity analysis and association with various selenium related traits.