Polymorphism of Bovine Lymphocyte Antigen DRB3.2 in Holstein Bulls of Iran Using PCR-RFLP

Authors

Maryam Parnian Department of Animal Science, Aboureyhan Campus, Tehran University, P.O. Box 11365-4117, Tehran, I,R. Iran and 2 National Institute for Genetic Engineering and Biotechnology, P.O. Box 14965-161, Tehran, I.R. Iran

Mohammad Reza Mollasalehi Animal Breeding Center, Karaj, I.R. Iran

Morteza Pashmi Department of Animal Science, Abhar Azad University, P.O. Box 22, Abhar, I.R. Iran and National Institute for Genetic Engineering and Biotechnology, P.O. Box 14965-161, Tehran, I.R. Iran

Seyed Ali Ghorashi National Institute for Genetic Engineering and Biotechnology, P.O. Box 14965-161, Tehran, I.R. Iran

Abstract:

The Holstein bulls (n=50) were genotyped for bovine lymphocyte antigen (BoLA-DRB3.2) alleles by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). Genomic DNA was extracted from bull semen using phenol-chloroform method. A two-step PCR was conducted in order to amplify a 284 base-pair fragment of the target gene. Amplicons were digested by RsaI, HaeIII and BstyI restriction endonuclease enzymes. Digested fragments were electrophoresed on 8% polyacrylamide gel and visualized after silver staining. Seventeen BoLA-DRB3.2 alleles were identified with frequencies ranging from 1 to 21%. Sixteen alleles were similar to those reported previously and one was a new allele which has not been reported before. The frequencies of alleles BoLA-DRB3.2 *3, *8, *10, *11, *12, *13, *15, *16, *21, *22, *23, *24, *28, *51, *iaa, *ibb, *qbb were 2, 9, 2, 14, 1, 2, 4, 10, 1, 14, 5, 21, 6, 6, 1, 1, and 1%, respectively. The seven most frequent alleles (BoLA-DRB3.2 *8, *11, *16, *22, *24, *28, *51) accounted for 80% of alleles in the investigated population. This data indicate that the BoLA-DRB3.2 locus is highly polymorphic in Holstein bulls of Iran.

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