شناسايي و تعيين الل هاي جديد در جايگاه ژني BoLA-DRB 3.2 گاوهاي سيستاني بر اساس روش هاي مولكولي

New alleles detection in BoLA-DRB3.2 locus of sistani cattle by molecular based typing method

This study was carried out for identification new alleles in BoLA-DRB3.2 locus of Sistani cattle. For mentioned purpose, bovine DNA was isolated from aliquots of 200 samples blood. Then amplification of the BoLA-DRB3 exon 2 was done by using Hemi Nested PCR, in two-step, and then followed by digestion with restriction endonucleases RsaI, HaeIII and BstX2I. As a whole, we identified 24 alleles in BoLA-DRB3.2 locus of Sistani cattle, that 21 of these alleles were similar to those reported in past studies. But the remaining 3 alleles including DRB3.2 *obc, *ibc, and *eac with respectively, 0.33%, 1.33% and 8.1% frequency, had not been identified in studies that carried out before. The obtained sequence of new patterns were submitted to the NCBI Gen Bank and registered with accession numbers as DQ486519, EU259858, EU259857 respectively. These results demonstrated that the BoLA DRB3.2 locus is highly polymorphic in Sistani cattle and the three new alleles, can be used as a selective index or breed marker in this species.