Open Access

Detection of the Bovine Leptin Gene rs29004488 Polymorphism by Introducing a New Restriction Enzyme Site For PCR-RFLP

1 Balıkesir University, Faculty of Veterinary Medicine, Department of Genetics, Balıkesir
2 Fırat University, Faculty of Veterinary Medicine, Department of Zootechny, Elazığ
3 Düzce University, Faculty of Medicine, Department of Medical Biology, Düzce
4 Fırat University Faculty of Medicine Department of Medical Biology, Elazığ

Abstract

Leptin is a hormone mainly synthesized in adipose tissue and plays an important role in the regulation of appetite, energy metabolism, body weight and reproductive functions. The rs29004488 polymorphism in the 2nd exon of the bovine leptin gene, which results in an amino acid change from arginine to cysteine, has been reported to be associated with economically important traits such as growth, carcass traits, milk yield and reproduction. In cattle, the leptin gene rs29004488 polymorphism can be detected by PCR-RFLP method using Kpn2I enzyme. However, this method has some limitations in terms of primer degradation. In this study, a new PCR-RFLP method was investigated to detect rs29004488 polymorphism of leptin gene in cattle. Using the created restriction site PCR (CRS-PCR) method, PstI enzyme recognition site was introduced into the PCR products, and rs29004488 polymorphism was successfully detected using newly designed primers and PstI enzyme. The obtained results were confirmed by DNA sequence analysis and comparison with Kpn2I enzyme cleavage results. This study demonstrates that the rs29004488 polymorphism can be detected by an efficient and low-cost method using CRS-PCR-RFLP and PstI enzyme.

 

Keywords

How to Cite

GURSES, M., BAYRAKTAR, M., YUCE, H., & ONALAN , E. (2023). Detection of the Bovine Leptin Gene rs29004488 Polymorphism by Introducing a New Restriction Enzyme Site For PCR-RFLP . MAS Journal of Applied Sciences, 8(Özel Sayı), 1024–1031. https://doi.org/10.5281/zenodo.10024106

References

📄 Buchanan, F.C., Fitzsimmons, C.J., Van Kessel, A.G., Thue, T.D., Winkelman-Sim, D.C., Schmutz, S.M., 2002. Association of a missense mutation in the bovine leptin gene with carcass fat content and leptin mRNA levels. Genetics Selection Evolution, 34(1): 105-116.
📄 Buchanan, F.C., Van Kessel, A.G., Waldner, C., Christensen, D.A., Laarveld, B., Schmutz, S.M., 2003. An association between a leptin single nucleotide polymorphism and milk and protein yield. Journal of Dairy Science. 86: 3164-3166.
📄 Chebel, R.C., Santos, J.E.P., 2011. Association between leptin single nucleotide polymorphism and reproductive performance of lactating Holstein cows. Animal Reproduction Science, 127(3-4): 126-134.
📄 Chessa, S., Nicolazzi, E.L., Nicoloso, L., Negrini, R., Marino, R., Vicario, D., Ajmone-Marsan, P., Valentini, A., Stefanon, B., 2015. Analysis of candidate SNPs affecting milk and functional traits in the dual-purpose Italian Simmental cattle. Livestock Science, 173: 1-8.
📄 Fathoni, A., Maharani, D., Aji, R.N., Choiri, R. and Sumadi, S., 2019. Polymorphism of the SNP g. 1180 C>T in leptin gene and its association with growth traits and linear body measurement in Kebumen Ongole Grade cattle. Journal of the Indonesian Tropical Animal Agriculture, 44(2): 125-134.