Detection of Lack of Function Mutation of P.R12X in SLC37A2 Locus in Montbeliarde and Holstein Cattle

The aim of this study was to identify the lack of function mutation of p.R12X affecting reproductive problems and abortion in SLC37A2 gene by PCR-SSCP and sequencing methods in Holstein and Montbeliarde cows, For this purpose, 250 blood samples (150 Holstein and 100 samples of Montbeliarde) were prepared and DNA was extracted by kit. A pair of specific primers was designed with Oligo7 software to amplify a fragment with length of 208 bp from exon 2 of the SLC37A2 gene, which contains mutation. Genotyping was done by SSCP method and allelic and genotypic frequencies and population genetic indices were calculated by POPGEN software. In SLC37A2 locus, two AA and AB genotypes with frequencies of 98.7 and 1.3, and 86 and 14% were observed in Holstein and Montbeliarde breeds, respectively. Also, SLC37A2 allelic frequencies in both Holstein and Montbeliarde for alleles A and B were estimated to be 99.3, 0.7 and 93 and 7%, respectively. The Fischerchr('39')s exact and Chi-square tests for comparing allelic and genotypic frequencies between two populations showed that there was a significant difference in allelic and genotypic frequencies between Holstein and Montbeliarde population (P <0.05). After genotyping, a sample of each genotype was sequenced in both end and the bioinformatics of the sequences obtained were performed with BioEdit software. The results showed that in the AB genotype of SLC37A2 gene in Holstein and Montbeliarde cows there was a lack of function mutation (p.R12X). Considering the role of p.R12X mutation in reproductive defects and abortion in Montbeliarde cattle, the results of this study can be directly used in the breeding programs to identify carrier animals and culling them from commercial and rural herds.