IDENTIFICATION OF HOLSTEIN COWS CARRIERS OF COMPLEX VERTEBRAL MALFORMATION BY HIGH RESOLUTION MELTING CURVES (HRM)

Andrea Branda Sica, Paula Nicolini, María Teresa Federici, Silvia Llambí

Resumo


The objective of this study was the optimization and implementation of a reliable and economical molecular screening method for the detection of the mutant allele of CVM (complex vertebral malformation, c.559G>T, SLC35A3) by HRM analysis, as well as analyzing its existence in a representative sample of Holstein cows from the Milk Genomic DNA Bank of Uruguay. The optimization of the HRM methodology in the RotorGene™ 6000 equipment (Corbett Life Science, Australia) by amplification of the 79 bp PCR products clearly differentiated the two genotypes: homozygous, wild type: GG; and heterozygous, carrier for the mutation CVM: GT (c.559G>T; SLC35A3). In the analyzed sample, the frequency of the mutant allele (T) for CVM was high (q = 0.032), with a prevalence of carrier cows of 6.45%. It is concluded that the PCR-HRM analysis is a fast, easily interpretable, low cost, and highly accurate technique for the detection of this mutation in Holstein cattle, which may be implemented in genetic selection programs.


Palavras-chave


Holstein; CVM; high-resolution dissociation curve.

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DOI: http://dx.doi.org/10.5380/avs.v24i4.65494