Genetic improvement has allowed great advances in selecting individuals with interest for determine characteristics. Knowledge about bovine genome and genetic markers along each chromosome allow the search for genetic variants that affect important productive and reproductive traits, making possible the genetic improvement. The goal of this study was to determinate the association between the POU1F1 gene with productive and reproductive traits in a population of Holstein cows from Antioquia, Colombia. The POU1F1 gene belongs to transcription factors POU, which regulates growth and development in animals. This gene participates on pituitary development and hormonal expression in mammals, regulating the production of growth hormone, thyroid stimulating hormone and prolactin. DNA was extracted from 523 samples, and 2 alleles were identified using the PCR-RFLP technique. The frequency for the alleles A and B was 21.26 and 78.74%, respectively, and the 3 genotypes AA, AB and BB showed frequencies of 2.87, 36.78, and 60.34%, respectively. The association between the genotypes and the studied traits was analyzed with a linear mixed model including the fixed effect of POU1F1 genotype, parity, year and month of birth, and the random effect of the animal. The analyzed SNP had no significant effect on the evaluated traits milk yield, protein percentage, fat percentage, calving interval and services per conception. The absence of association between the SNP and evaluated traits could be due to 2 reasons; the interaction of this gene with other genes involved in the POU1F1 pathway which may hide the individual effect of this SNP on the evaluated traits, and the genotype-environment interaction, the latter has a large effect on gene expression. This way, the POU1F1 gene is not a good candidate to be used in breeding programs assisted by molecular markers (MAS) for the evaluated population.