Keragaman Gen CYP26B1 Daerah CpG Island pada Sapi Bali, PO, dan Limousin Menggunakan Metode PCR-RFLP

Abstract

Penelitian ini bertujuan mengidentifikasi keragaman genetik gen CYP26B1 daerah CpG island pada tiga bangsa sapi (bali, PO, dan limousin) menggunakan metode polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP). Jumlah sampel yang digunakan 53 sampel sapi meliputi: 30 sapi bali, 13 sapi PO, dan 10 sapi limousin. Keragaman gen CYP26B1 dilakukan menggunakan metode PCR-RFLP dengan enzim AciI. Hasil pemotongan fragmen DNA dengan enzim restriksi AciI menunjukkan adanya polimorfisme yang menghasilkan tiga genotipe, yaitu TT, CT, dan CC. Alel T ditemukan dominan pada populasi sapi bali (0,92), limousin (0,55), dan PO (0,50), sedangkan alel C sebagai alel minor. Analisis statistik mengonfirmasi bahwa frekuensi genotipe pada ketiga bangsa sapi berada dalam kesetimbangan Hardy-Weinberg. Melalui penyejajaran sekuens, terdeteksi mutasi transisi T>C serta delesi yang keduanya terletak pada CpG island di daerah promotor. Meskipun mutasi transisi bersifat silent, keberadaan variasi genetik pada wilayah ini berpotensi memodulasi tingkat ekspresi gen.

This study aims to identify the genetic diversity of the CYP26B1 gene in the CpG island region across three cattle breeds (bali, PO, and limousin) using the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method. A total of 53 cattle samples were utilized comparising 30 bali cattle, 13 PO cattle, and 10 limousin cattle. The diversity of the CYP26B1 gene was analyzed via the PCR-RFLP method using the AciI enzyme. DNA fragment digestion with the AciI restriction enzyme revealed polymorphisms resulting in three genotypes: TT, CT, and CC. The T allele was found to be dominant in bali (0.92), limousin (0.55), and PO (0.50) cattle populations, while the C allele served as the minor allele. Statistical analysis confirmed that the genotype frequencies in the three cattle breeds were in Hardy-Weinberg equilibrium. Sequence alignment detected a T>C transition mutation and deletion, both located within the CpG island of the promoter region. Although the transition mutation is silent, the presence of genetic variation in this region has potential to modulates gene expression levels.