The bovin leukocyte antigen (BoLA) system is the major histocompatibility complex (MHC) of cattle. MHC genes encode highly polyorphic cell-surface molecules which present antigenic peptides to T-cell, thereby playing an essential role in immune response to foreign agents and the differentiation among the species. The BoLA-DR region contains at least three DRB genes but only the DRB3 gene is expressed at a high level in peripheral blood lymphocytes. In this study, we analyzed the genetic polymorphism of the BoLA DRB3 exon2 gene from 20 Hanwoo (Korean native cattle) and 21 Holstein by polymerase chain reaction combined with restriction fragment length polymorphism (PCR-RELP) with Rsal, BstYI and HaeIII. In the 42 alleles of Holstein, DRB3*0101 (21.4%) and *1501 (11.9%) were major genotypes, whereas DRB3*1401 (20%) and *2001 (10%) were major genotypes in 40 alleles of Hanwoo. We are found two new allele types of Hanwoo, which were named with DRB3kor1 (CenBank accession number AF008234) and DRB3kor2 (AF008235), using DNA sequence analysis.