Polymorphism of the myostatin gene exon 1 using PCR-RFLP technique in five beef cattle in Indonesia

Abstract

This study aims to analyze the polymorphism of the myostatin (MSTN) gene in exon 1 at SNPc.111G>C (rs523392653) and SNPc.267G>A (rs383271508) on various beef cattle in Indonesia. A total of 136 DNA samples were analyzed in this study, consisting of 33 heads of Madura, 31 heads of PO, 36 heads of Bali, 18 heads of Simmental, and 18 heads of Limousin cattle. The DNA sample was collected from the Animal Molecular Genetic Laboratory, Faculty of Animal Science, IPB University. Along 608 bp of DNA sequence was amplified using PCR with forward primer was 5′-CAA GTT GTC TCT CAG ACT GG-3′ and reverse primer was 5′-CTC CTC CTT ACA TAC AAG CC-3′ and annealing temperature was 59°C. The amplified DNA was then digested with HaeIII and AluI restriction enzymes using the RFLP method to determine polymorphism. Genotype frequencies, allele frequencies, PIC, and heterozygosity values were calculated with the PopGen32 program. The HaeIII enzyme produces two genotypes, GG and CG, while the AluI enzyme produces three: GG, AG, and AA. SNPc.111G>C polymorphism was found in Madura and PO cattle, while SNPc.267G>A polymorphism was found in Madura, PO, and Bali cattle. Both Simmental and Limousin cattle were monomorphic.