ASSOCIATION OF AN INDEL FROM THE REGULATORY REGION OF THE MELATONIN RECEPTOR GENE (MTRN1A) WITH MILK PERFORMANCE OF MESTIZED BUFFALOES IN THE AMAZON REGION
Buffalo farming, PCR, Genetic improvement, Indel.
Buffalo farming is an important agricultural activity, especially in developing countries, being used for traction, meat and milk production. In the Amazon Region, these animals have good productive and reproductive performance, even when raised in adverse conditions. Animal genetic improvement programs aim to multiply individuals with desirable characteristics related to production and reproduction. Therefore, it is essential to identify molecular markers relevant to phenotypes of economic importance. The present study aims to associate a deletion block (INDEL) in the promoter region of the melatonin 1A receptor gene (MTRN1A) with zootechnical data on milk production in a herd of crossbred buffaloes from the Amazon region. Blood samples will be collected from 85 buffaloes from a property located in the municipality of Bujaru, state of Pará. Afterwards, DNA extraction will be carried out using the phenolic method. The genomic DNA will have its specific regions from the primers, amplified by PCR (Polymerase Chain Reaction), with a final volume of 25 µL. All PCR products will be purified using a commercial kit following its recommendations and subjected to sequencing using the Sanger method. The sequences obtained will be edited, compared with buffalo reference sequences in NCBI GenBank and aligned for analysis. The sequencing results obtained will be tabulated and submitted to the Genepop v.5 program, which will determine the allelic and genotypic frequencies, observed and expected heterozygosity, coefficients of inbreeding (FIS), Hardy-Weinberg deviation and the probability of Hardy-Weinberg equilibrium (P> 0.05). The associations will be carried out by ANOVA analysis with a fixed effect of the genotype and mean comparison using the Tukey test, using the SAS OnDmand program, with a significance level of 0.05%.