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Asian Journal of Animal Sciences

Year: 2017 | Volume: 11 | Issue: 1 | Page No.: 40-46
Research Article

Sequence Analysis and Identification of Allele Distribution of Melanocortin 1 Receptor (MC1R) Gene in Indonesian Cattle (Bos sondaicus×Bos indicus)

Tety Hartatik
Laboratory of Animal Genetic and Breeding, Department of Animal Breeding and Reproduction, Faculty of Animal Science, Universitas Gadjah Mada, 55281 Yogyakarta, Indonesia
Background and Objective: The melanocortin-1-receptor (MC1R) gene is an important candidate gene for the coat color trait. Since, Indonesia has the diversity of local cattle with various type of colour, it is very interest to investigate the sequence of MC1R gene. The aim of this study was to identify polymorphism of melanocortin-1-receptor gene in Indonesian cattle based on sequence analysis. This study was conducted at Laboratory of Animal Genetic and Breeding, Department of Animal Breeding and Reproduction, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta, between August, 2012 and December, 2016. Materials and Methods: The total number of 164 cattle were used in this study that consist of Aceh cattle (n = 25), Filial Ongole cattle (n = 6), Limousin×Madura (Limura, n = 11), Madura cattle (n = 7), Brahman cattle (n = 60), Pesisir cattle (n = 9) and Bali cattle from Kupang (n = 46). A fragment of 296 base pair of melanocortin 1 receptor (MC1R) gene was amplified by forward primer: 5’-GGA CCC TGA GAG CAA GCA C-3’, reverse primer: 5’-CTC ACC TTC AGG GAT GGT CTA-3’. Polymerase Chain Reaction (PCR) products were digested with MspI enzyme for identifying of restriction pattern (PCR-RFLP result). Sequencing of the representative sample was conducted base on PCR-RFLP result with different type of polymorphism. Results: The PCR-RFLP analysis showed three pattern of restriction, there are EE genotype, Ee genotype and undigested sample with ee genotype. Sequence analysis show point mutation along the 296 bp of MC1R gene with two type of mutation (substitution 147C→T and deletion/insertion 162indelG). Bali cattle (Bos soandaicus ) and Aceh cattle (Bos indicus) revealed the monomorphic with EE genotype. The others (PO, Limura, Madura, Pesisir and Brahman) showed the polymorphic with equilibrium in chi-square analysis except Limura (Bos soandaicus×Bos indicus×Bos taurus ) with χ2 = 7.64 for Limura population. Conclusion: Local cattle such as Bali and Aceh cattle have higher E allele distribution compare to that of the other local cattle. Indeed, the homozygosity of MC1R gene (EE) in local cattle also higher than that of the crossbred cattle. The MC1R gene can be used to identify the genetic variation of local cattle and showed heterogeneity in crossed breed cattle.
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