ABSTRACT
O objetivo desta pesquisa foi avaliar os SNPs rs471462296, rs456245081 e rs438495570 do gene DGAT1 em bovinos Nelore. Foram analisados 109 bovinos. A extração do DNA genômico foi realizada do sangue dos animais, usando-se o kit Ilustra Blood Genomic Prep Mini Spin® (GE Healthcare, UK). A concentração e o grau de pureza do DNA foram determinados por meio de espectrofotômetro (Nanodrop - Thermo Fisher Scientifc, USA). A genotipagem dos SNPs ocorreu mediante o emprego do ensaio Taqman® (Applied Biosystems, USA). Na análise genômica, não foram encontradas alterações nas frequências alélicas e genotípicas (P≥0,05) para os SNPs testados. Dessa forma, a região 5'UTR analisada apresentou-se monomórfica e a variação de SNPs não foi observada, o que limita seu uso como marcadores moleculares para o gene DGAT1 em Nelore.(AU)
Subject(s)
Animals , Cattle , Cattle/genetics , Phenotype , GenotypeABSTRACT
A mixture of bovine DNA from a male and a female Jersey (Bos taurus taurus) bred in different proportions was used to determine the sensitivity of PCR to amplify and discriminate the bovine DNA samples. Samples were obtained from the peripheral blood of a bull and a heifer and DNA was isolated using a commercial kit for extraction and purification of nucleic acids. Two primers sets were designed to flank genomic regions: one autosomal and one Y-specific. DNA samples were diluted in water to a final concentration of 4x10-14 ng. The results showed positive amplification of the samples diluted to a concentration of 4x10-10ng and 4x10-4ng for the autosomal and Y-specific regions, respectively. PCR was able to discriminate the male DNA in a mixture of 99:1 (DNA ♀: DNA ♂) heifer to bull ratio. Therefore, the PCR was successful in amplifying the bovine genome in samples containing low concentrations of DNA. Thus, PCR can be used as a sensitive and efficient tool to determine the sex of the fetus in pregnant cows, helping to promote correct and efficient animal management, sex selection, and breeding in commercial herds.