Review

Identification of Single Nucleotide Polymorphism and Restriction Enzyme in FASN and CAPN-1 Genes in Ongole Grade cattle

Rahmatika Choiria1    Rusman2   Dyah Maharani1,*   

1Department of Animal Breeding and Reproduction, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
2Department of Animal Product and Technology, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia

Abstract

FASN have responsible for the activity of FASN enzymes and fatty acid synthesis. CAPN-1 gene is the primary enzyme in the postmortem tenderization process. This study aims to identify Single Nucleotide Polymorphism (SNP) FASN and CAPN-1 genes, and restriction enzymes that will be used in determining animal genotype. Thirty-eight meat samples from 22 Ongole Grade cattle slaughter in Giwangan slaughter house and 16 Kebumen Ongole Grade cattle were collected in Kebumen slaughter house. The samples were analyzed using the PCR method and sequencing for SNP identification. Two SNPs of FASN were detected in intron region (g.17066 G>A and g.17104 T>C) by direct sequencing. Restriction enzyme FatI has been identified to digest FASN gene target in the SNP g.17104T>C. However, no enzyme detected in SNP g.17066 G>A of FASN gene. Nine SNPs was detected with GenBank’s alignments (g.16838G>A, g.16844G>C, g.16986G>A, g.17058C>A, g.17091G>A, g.17100G>A, g.17104T>C, g.17194C>A, g.17195C>A). Using direct sequencing, the CAPN-1 gene was revealed three synonymous polymorphism (g.4481A>G, g.4519C>A, which located in intron 4 and g.4631T>C which located in intron 5) and one non-synonymous polymorphism (g.4554C>T) located in exon 4. Only one SNP detected (g.4732T>G) with GenBank’s alignment. The AciI and FauI for SNP g.4481A>G, MnlI and BccI for SNP g.4519C>A, and Sau3AI, MboI, DpnII, DpnI, and BclI restriction enzyme could digest the SNP g.4554C>T. In conclusion, the identified the SNPs could be utilized for genotyping of Ongole Grade and Kebumen Ongole Grade cattle for future studies

Figures & Tables

Figure 1.Multiple alignment of FASN gene with 4 GenBank Acc No. NC_03228, AF285607, and NC_037346