Analisis Keragaman SNP g.1117G>A Gen HSP70 pada Sapi Limousine, Belgian Blue, Wagyu, dan FH Menggunakan Metode PCR-RFLP

Abstract

Gen HSP70 merupakan salah satu gen yang berfungsi untuk mengatasi cekaman panas. Penelitian ini bertujuan untuk menganalisis keragaman gen HSP70 menggunakan metode PCR-RFLP dengan ezim pemotong BanI. Sampel DNA yang digunakan sebanyak 79 sampel sapi Bos taurus yang terdiri dari 58 ekor tipe pedaging (limousine, belgian blue, wagyu) dan 21 ekor tipe perah (FH). Keragaman genetik gen HSP70 dihitung frekuensi genotipe dan alel, nilai heterozigositas, serta keseimbangan Hardy-Weinberg menggunakan program PopGen32. Amplifikasi fragmen gen HSP70 pada daerah 5’UTR dengan suhu annealing 61 oC selama 15 detik menghasilkan panjang produk PCR sebesar 539 bp dan diperoleh 2 macam alel (A dan G) dan 3 macam genotipe (GG, AG, AA). Panjang produk genotipe yang dihasilkan yaitu GG (369 bp dan 106 bp), AG (475 bp, 369 bp, dan 106 bp), dan AA (475 bp). Hasil menunjukkan bahwa keragaman gen HSP70 pada sapi Bos taurus tipe pedaging dan perah bersifat polimorfik dan berada dalam keseimbangan Hardy-Weinberg.

The HSP70 gene is one of the genes that functions to overcome heat stress. This study aimed to analyze the diversity of the HSP70 gene using the PCR-RFLP method with the BanI cutting enzyme. The DNA samples used were 79 samples of Bos taurus cattle consisting of 58 beef breeds (limousine, belgian blue, wagyu) and 21 dairy cows (FH). The genetic diversity of the HSP70 gene was calculated for genotype and allele frequencies, heterozygosity values, and Hardy-Weinberg balance using the PopGen32 program. Amplification of the HSP70 gene fragment in the 5'UTR region with annealing temperature of 61 oC for 15 seconds resulted in a PCR product length of 539 bp and 2 alleles (A and G) and 3 genotypes (GG, AG, AA) were obtained. The length of the genotype products produced were GG (369 bp and 106 bp), AG (475 bp, 369 bp, and 106 bp), and AA (475 bp). The results showed that the diversity of the HSP70 gene in beef and dairy Bos taurus cattle was polymorphic and was in Hardy-Weinberg equilibrium.