Judul: Genetic Diversity of Promoter and 5’ Untranslated Region of HSP70 Gene in Beef Cattle
Abstrak: Indonesia memiliki beragam jenis sapi baik asli, lokal, maupun eksotik yang berpotensi sebagai sumber protein hewani. Pemanasan global telah menjadi ancaman bagi produktivitas ternak yang berdampak negatif pada pertumbuhan, reproduksi, produksi susu, dan sifat karkas ternak. Adaptasi adalah mekanisme penting bagi ternak jika pemicu stres lingkungan berlangsung dalam kurun waktu yang lama. Salah satu mekanisme adaptasi adalah heat shock protein, termasuk HSP70 (heat shock protein 70) yang merupakan biomarker penting dalam mekanisme biokimia terhadap stres panas. Penelitian ini bertujuan untuk mengidentifikasi keragaman daerah promotor dan 5' UTR (untranslated region) gen HSP70 pada sapi pedaging di Indonesia. Total sampel yang digunakan adalah sebanyak 86 sampel darah yang terdiri dari sapi bali, madura, PO (peranakan ongole), limosin, dan belgian blue. DNA Sampel selanjutnya dianalisis menggunakan metode direct sequencing. Hasil genotyping menunjukkan adanya lima SNP (single nucleotide polymorphism) di daerah promotor (g.-393T>C, g.- 343C>A, g.-202T>C, g.-69T>G) dan tiga SNP di 5' UTR (g.19A>G, g.45C>T, g.100_101ins31). Frekuensi alel yang ditemukan pada sapi PO, limosin dan belgian blue berada dalam keseimbangan Hardy-Weinberg, sedangkan pada populasi sapi bali dan sapi madura frekuensi alelnya disequilibrium. Secara umum keragaman alel yang diamati berkisar rendah hingga tinggi (0.26-1.00), SNP g.19A>G merupakan SNP yang memiliki keragaman tertinggi. Ditemukan bahwa SNP g.-69T>G berikatan dengan faktor transkripsi dari NF-Y dan CAAT box. Selain itu, ditemukan insersi 30 pb (pasang basa) (g.100_101ins31) pada sapi bali dan sapi madura yang belum pernah dilaporkan sebelumnya., Global warming has become a threat of livestock productivity by negatively affecting growth, reproduction, milk yield, and carcass traits of the livestock. Adaptation is a crucial mechanism for animals if the environmental stressor is present for an extended period. One of the mechanisms regulations of adaptation is Heat shock protein, include HSP70 (Heat Shock Protein 70), which is an essential biomarker as a biochemical mechanism against heat shock. This study aims to identify genetic diversity in the promoter area and 5' UTR (untranslated region) HSP70) gene in several beef cattle in Indonesia. A total of 86 blood samples of bali, madura, PO (peranakan ongole), limousine, and BB (belgian blue) cattle were used in this study. The extracted DNA of all blood samples was then analyzed using the direct sequencing method. The genotyping results showed the presence of five SNP (Single Nucleotide Polymorphism) in the promoter region (g.-393T>C, g.-343C>A, g.-202T>C and g.-69T>G) and three SNPs at 5' UTR (g.19A>G, g.45C>T, and g.100_101ins31). The frequency of SNP alleles found in PO, limousine, and BB cattle was in equilibrium, but in bali and madura cattle populations, the allele frequency was disequilibrium. In general, the level of diversity of observed alleles ranged from low to high (0.26-1.00), where SNP g.19A>G had the highest diversity. It was successfully revealed in this analysis that the SNP g.-69T > G binds to both the NF-Y and CAAT box transcription factor. In addition, the 30 bp (base pair) insertions (g.100_101ins31) that were identified in bali and madura cattle have never been reported in previous research studies.
Keyword: 5’-UTR, HSP70, promotor, sapi pedaging, SNP