SNP g.1117G>A HSP70 Gene Polymorphism in Domestic Beef Cattle using PCR-RFLP Method

Abstract

Temperature increases have detrimental impacts on livestock, such as decreased metabolism, reproduction, and performance. The response of livestock to heat stress can be shown by the expression of HSP70 gene. This study aims to identify the SNP g.1117G>A HSP70 gene polymorphism (HSP70|BanI) using the PCR-RFLP method. This study used 118 DNA samples of 5 breeds of cattle consisting of 20 heads of PO cattle, 20 heads of Madura cattle, 12 heads of Kuantan cattle, 53 heads of Bali cattle, and 13 heads of Pesisir cattle. The PCR-RFLP approach was used to analyze the diversity of the HSP70 gene. The enzyme used for cutting was BanI with forwarding primer 5‘-GAA AGG AAA AAA GAG ACA GA-3’ and reverse primers 5‘-AAT ACG CAG GAG TAG GTG GT-3’. The variation of the HSP gene was analyzed using allele frequency, genotype frequency, and level of heterozygosity. The Hardy-Weinberg equilibrium was evaluated using the Chi-Square test on PopGen 32. HSP70 gene amplification resulted in 539 bp fragments and 2 alleles (A and G). The HSP70 gene consisted of three genotypes i.e., AA (475 bp), AG (475 bp, 369 bp, and 106 bp), and GG (369 bp and 106 bp). The highest allele frequency of the A allele was found in PO and G allele in Bali cattle. Based on the results, it can be concluded that the HSP70 gene in 5’ UTR (untranslated region) in local beef cattle was polymorphic.