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Journal of Dairy Science Vol. 85 No. 6 1630-1632
© 2002 by American Dairy Science Association ®
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Short Communication: Characterization of DRB3 Alleles in the MHC of Japanese Shorthorn Cattle by Polymerase Chain Reaction-Sequence-Based Typing

S. Takeshima 1, Y. Nakai 2, M. Ohta 2, and Y. Aida 1

1 Retrovirus Research Unit, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0114, Japan Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidoriamamiya-machi Aoba-ku, Sendai 981-8555, Japan
2 Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidoriamamiya-machi Aoba-ku, Sendai 981-8555, Japan

A study was made of exon 2 of the bovine leukocyte antigen BoLA-DRB3 gene of 176 Japanese Shorthorn cattle at six farms in Japan using polymerase chain reaction-sequence-based typing (PCR-SBT). An initial round of PCR using conserved locus-specific primers, a second round of PCR using a locus-specific primer, and at least one sequence-specific primer (SSP), followed by direct sequencing of products of PCR with SSP were conducted. Twenty-one BoLA-DRB3 alleles were identified with frequencies ranging from 0.3 to 19.6% in 176 individuals, and two of these alleles were new alleles that have not been reported previously. The three most frequently observed alleles (DRB3*1201, *0301, and *0801) accounted for 43.8% of the alleles in the population of these herds. Next, we tested the products of amplification by PCR of BoLA-DRB3 exon 2 with RsaI, BstYI, and HaeIII, and identified 18 previously described PCR-restriction fragment length polymorphism (RFLP) types. The PCR-RFLP types reflected the results of PCR-SBT exactly. Our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Japanese Shorthorn cattle.

Key Words: BoLA-DRB3 exon 2 • PCR-sequence-based typing • genetic polymorphism • Japanese Shorthorn cattle

Submitted on November 26, 2001
Accepted on January 7, 2002




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