Detection of Lack of Function Mutation of p.Q879X Affecting Abortion in APAF1 Gene in Holstein Cattle

Today, reproductive problems are one of the major concerns in the dairy industry. The aim of this study was to detect the lack of function mutation of p.Q579X affecting spontaneous abortion and reproduction problems in APAF1 gene in Holstein cattle. In total, 200 blood samples were prepared and DNA was extracted with commercial kit. Then a specific pair of primers was designed to amplify a fragment with lengths of 456 bp from the exons 11 of the APAF1 gene and genotyping was performed by PCR-SSCP method. In APAF1 locus two alleles of A and B and two genotypes of AA and AB with frequencies of 96.76, 3.24 and 93.51 and 6.49 were observed, respectively. The BB genotype was not observed in the studied samples. After determining the genotype of the samples, two samples from each genotype were sequenced from both end and alignment of the obtained sequences and their comparison with the reference sequence from the gene bank to detect the mutation was carried out using BioEdit (version7.0.9.0) software. The results of bioinformatics analysis showed that the lack of function mutation of p.Q579X, which is one of the mutations affecting abortion in Holstein cattle, is present in the AB genotype of APAF1 gene in Holstein cattle in Iran. Given that this mutation causes abortion in cow in homozygous form, the results of this study can be directly used in breeding programs in order to identify the carriers of this mutation and culling them from Holstein dairy herd.