Document Type: Original Article

Authors

Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

Abstract

Objective: Diacylglycerol acyltransferase 1 is a microsomal enzyme that catalyzes the final step of triglyceride synthesis. The objective of this project is to check out the polymorphism at the exon 8 region of DGAT1 gene using PCR-SSCP technique in Iranian water buffaloes (Bubalus bubalis). Recent activities have shown that a significant association between lysine at amino acid position 232 with higher milk fat content, whereas an alanine at this position is marker data of lower milk fat content and higher milk yield. Methods: In this study we analyzed buffaloes from 5 provinces of Iran. Three PCR-SSCP patterns were found. Results: DNA sequencing analysis showed that all of the patterns had the motif “AA” at position 6962-6963 (K allele) encoding lysine, which is similar to “AA” at position 10433-10434 in Bos indicus and also different buffalo breeds at position 6962-6963. Therefore exon 8 is a conservative region in Bos indicus and all buffalo breeds which is associated with higher milk fat content. Moreover, we found a base transition at position 7036 C/T and we have heterozygosity of both “C and T”, which is located at intron 8 of DGAT1 gene in few samples. Conclusions: this transition is a nonsense mutation which causes no change in amino acid sequence. This mutation has occurred at the intron 8 of DGAT1 gene in Iranian buffaloes.

Keywords

Bassam B. J., Caetano-Anolles, G. and Gresshoff, P .M. 1991. Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal Biochem. 196:80-83.
Buraczynska, M., Ksiazek, P., Zaluska, W., Nowicka, T. and Ksiazek, A. 2004. Endothelial nitric oxide synthase gene intron 4 polymorphism in patients with end-stage renal disease. Neph Dial Transp. 19:2302-2306.
Cases, S., Smith, S. J., Zheng, Y. W., Myers, H. M., Lear, S. R., et al. 1998. Identification of a gene encoding an acyl CoA: Diacylglycerol acyltransferase, a key enzyme in triacylglycerol synthesis. Proc Natl Acad Sci.
95(22):13018-13023.
Coppieters, W. J., Riquet, J. J., Arranz, P., Berzi, N. and Cambisano N. 1998. A QTL with major effect on milk yield and composition maps to bovine chromosome 14. Mamm. Genome. 9:540-544.
Furbass, R., Winter, A., Fries, F., Kuhn, C. 2006. Alleles of the bovine DGAT1 variable number of tandem repeat associated with a milk fat QTL at chromosome 14 can stimulate gene expression. Physiol Genomics. 25:116120.
Grisart, B., Coppieters, W., Farnir, F., Karim, L., Ford, C., et al. 2002. Positional candidate cloning of a QTL in dairy cattle: idenrification of amissense mutation in the bovine DGAT1 gene with major effect on milk yield composition.
Genome Res. 12: 222-231.
Grisart, B., Farnir, F., Karim, L., Cambisano, N., Kim J. J., et al. 2004. Genetic and functional confirmation of the causality of the DGAT1 K232A quantitative trait nucleotide in affecting milk yield and composition. Proc Natl Acad Sci USA. 101:2398-2403.
Gruszczynska, J., Brokowska, K., Charon, K. M. and Swiderek, W. P. 2005. Restriction fragment length polymorphism of exon 2 Ovar-DRB1 gene in Polish Heath Sheep and Polish Lowland Sheep. J Appl Genet. 46:311-314.
Hastings, M.L. and A.R. Krainer, 2001. Pre-mRNA splicing in the new millennium. Curr Opin Cell Biol. 13:302-209.
Kaupe, B., Winter, A., Fries, R., Erhard, G. 2004. DGAT1 polymorphism in Bos indicus and Bos taurus cattle breeds. J Dairy Res. 71:182-187.
Lacorte, G. A., Machado, M. A., Martinez, M. L., Campos, A. L., Maciel, R. P. et al. 2006. DGAT1 K232A polymorphism in Brazilian cattle breeds. Genet Mol Res. 5:475-482.
Miller, S. A., Dykes, D. D., Polesky, H. F. 1988. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research. 16:12-15.
Mishra, B., Tantia, M. S., Kumar, S. T. B., Vijh, R. K. 2007. Characterization of the DGAT1gene in the Indian buffalo (Bubalus bubalis). Genet Mol Biol. 30:1097-1100.
Raut, A. A., Kumar, A., Kala, S. N., Chhokar, V., Rana, N., et al. 2012. Identification of novel single nucleotide polymorphisms in the DGAT1 gene of the buffaloes by PCR-SSCP. Genet and Molec Biol. 35.3:610-613.
Ripoli, M. V., Corva, P., Giovambattista, G. 2006. Analysis of a polymorphism in the DGAT1 gene in 14 cattle breeds through PCR-SSCP methods. Res Vet Sci. 80:287-290.
Smith, S. J., Cases, S., Jensen, D. R., Chen, H. C., Sande, E. et al. 2000. Obesity resistance and multiple mechanisms of triglyceride synthesis in mice lacking DGAT. Nat Genet. 25:87-90.
Spelman, R. J., Ford, C. A., McElhinney, P., Gregory, G. C., Snell, R. G. 2002. Characterization of the DGAT1 gene in the NewZealand dairy population. J Dairy Sci. 85:3514- 3517.
Tantia, M. S., Vijh, R. K., Mishra, B. P., Mishra, B., Kumar, B. S. T., et al. 2006. DGAT1 and ABCG2 polymorphism in Indian cattle (Bos indicus) and buffalo (Bubalus bubalis) breeds. BMC Vet Res 2:e32.
Thaller, G., Kuhn, C., Winter, A. 2003. DGAT1, a new positional and functional candidate gene for intramascular fat deposition in cattle. Anim Genet.
34:354-357.
Venkatachalapathy, R. T., Sharma, A., Battacharta, T. K., Sukla, S. 2013. Single nucleotide polymorphism in DGAT1 locus of Indian cattle and buffalo breeds. Anim Genet Divi. (6).3:68-71.
Weller, J. I., Golik, M., Seroussi, E., Ezra, E., Ron, M. 2003. Population-wide analysis of a QTL affecting milk-fat production in the Israelian Holstein population. J Dairy Sci. 86:2219-2227.
Winter, A., Kramer, W., Werner, F., Kollers, S., Kata, S., et al. 2002. Association of a lysine/232/alanine polymorphism in a bovine gene encoding acyl-CoA:
Diacylglycerol acyltransferase (DGAT1) with variation at a quantitative trait locus for milk fat content. Proc Natl Acad Sci USA. 99:9300-9305.
Yuan, J., Zhou, J., Deng, X., Hu, X., Li, N. 2007. Molecular cloning and single nucleotide polymorphism detection of buffalo DGAT1 gene. Biochem Genet. 45:611-621.
Zhang, Q., Biochard, D., Hoeschele, I., Ernst, C., Eggen, A., et al. 1998. Mapping quantitative trait loci for milk production and health of dairy cattle in a large outbred pedigree. Genetics. 149:1950-1973.