Polymorphism of the DLA-DRB1 exon 2 gene in dogs analyzed by PCR-RFLP and PCR-SSCP

IT V; VILLEGAS CASTAGNASSO EE; DIAZ S; CASTELLANO MC; GIOVAMBATTISTA G

Journal of Zhejiang University

Año: 2008

The polymorphism of the canine major histocompatibility complex class II DRB1 gene 4 was analyzed. Genomic DNA was extracted from 80 dogs and the exon 2 of DLA5was analyzed. Genomic DNA was extracted from 80 dogs and the exon 2 of DLA5 DRB1 gene was analysed by PCR-RFLP and PCR-SSCP methods. Fifty four DLA66 DRB1 exon 2 previously reported sequences were analysed for HaeIII, RsaI and MspI 7 restriction sites. In the screened dog sample, the PCR-RFLP analysis revealed eightrestriction sites. In the screened dog sample, the PCR-RFLP analysis revealed eight 8 HaeIII patterns, six RsaI patterns and four MspI patterns of the thirty three previouslyHaeIII patterns, six RsaI patterns and four MspI patterns of the thirty three previously 9 defined alleles .The PCR-SSCP analysis exhibited thirteen different band patterns. Ourdefined alleles .The PCR-SSCP analysis exhibited thirteen different band patterns. Our 10 results suggested that SSCP method is more accurate to detect DLA polymorphism thanresults suggested that SSCP method is more accurate to detect DLA polymorphism than 11 RFLP technique. The improvement of both should provide a simple and rapid techniqueRFLP technique. The improvement of both should provide a simple and rapid technique 12 for DLA-DRB1 typing in dogs. In the future this will be useful in the development offor DLA-DRB1 typing in dogs. In the future this will be useful in the development of 13 strategies to study the association between the MHC and many dog diseases.strategies to study the association between the MHC and many dog diseases.