Gaur-catlle hybrids had been reported and they were often carrying intermediate characters genetically and phenotipically of their parents. In this study, 702-bp intron 5 PIT-1 gene, the functional gene of bovine genome, was sequenced and analyzed to compare and evaluate the genetic variation between local cattle (BN), hybrid calve (BL) and wild gaur (BT). Aligment of 702 bp sequence of PIT-1 gene from those samples and reference sequences from Genbank showed 14 mutations (substitution, deletion, insertion). All mutations were observed both in wild gaur Bos gaurus (BT-B and BT-C) and hybrid calves (BL-1, BL-2 and BL-3), not in domestic cattle (local cattle and both B. taurus and B. indicus). Our study revealed a genetic envidence for hybridization of wild gaur Bos gaurus and local cattle Bos taurus by nucler DNA of PIT-1 gene sequence. Keywords: Gaur, hybrid calve, local cattle, PIT-1 gene sequence.