Study of mutations available in first-halfexon 2 of GDF9 gene in crossbred sheep born from crossing of Romanov rams with Kermani ewes

Document Type : Genetics & breeding

Authors

1 Shahid Bahonar University of Kerman

2 Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran

3 Department of Animal Science, Faculty of Animal and Agricultural Sciences Engineering, Faculty of Agriculture and Natural Resources, Karaj, University of Tehran

Abstract

For decreasing the numberofbreedingewes onpastures and prevention of demolition pastures, Animal breedingprograms are necessary on genes with major effects on litter size in Iranian sheep breeds for Identify effective candidate genes on these economical traits .The GDF9 gene is one of the most important effective factors on litter size in sheep. The aim of the present study was to identify G2, G3 and G4 mutations in exon 2 of GDF9 gene in crossbred sheep (Romanov rams×Kermani ewes) using PCR-SSCP. Blood samples were collected from jugular vein of 121 crossbred individuals. Genomic DNA was extracted using salting-out method. Partial region of exon 2 (633 bp segments) of GDF9 gene was amplified with designed specific primers. The single stranded conformation polymorphism (SSCP) patterns of PCR products were studied using acrylamide gel electrophoresis and silver-nitrate staining method. Finally, we obtained 5 banding patterns 1, 2, 3, 4 and 5 with frequencies of 0.314, 0.024, 0.289, 0.232 & 0.141, respectively. The sequencing results showed presence 5 mutations in the studied population.

Keywords


1- Badbarin, N., S.Z.A. Mirhoseini., A. Bahmani., and R. seidsharifi. 2013. Study polymorphism the Growth Differentiation Factor 9(GDF9) gene and correlation it with weight quality in gharehghol sheep. Animal and birds congress,north Iran,1073-1076. (In Persian)
2- Bahmani, A., S.Z.A. Mirhoseini., B. Dlirsafat., z.Ansari. 2010. Study polymorphism the Growth Differentiation Factor 9 (GDF9) gene in gharehgholsheep using PCR-SSCP. In: Proceedings of 4th Animal science congress,Iran3590-3593. (InPersian)
3- Barker, J. S. F. 2001. Conservation and management of genetic diversity: a domestic animal perspective .Canadian Journal of Forest Research. 31:588-595.
4- Barzegari, A., S. Atashpaz., K. Ghabili., Z. Nemati., M. Rustaei., and R. Azarbaijani. 2010. PolymorpHisms in GDF9 and BMP15 associated with fertility and ovulation rate in Moghani and Ghezel sheep in Iran. Reproduction in Domestic Animals. 45: 666–669.
5- Chu, M. X., G.H. Cheng., L. Chen, Fang., and S.C. Ye. 2005. Study on morphogenetic protein 15 as a candidate gene for prolificacy of Small Tailed Han sheep and Hu sheep. Journal Anhui Agriculture University,32:278-282.
6- Dong, J., D.F. Altertini, K. Nishimori, T. Rajendra Kumar, N. Lu and M.M. Matzuk. 1996. Growth ifferentiation factor-9 is required during early ovarian folliculogenesis. Nature Genetics, 383:531-535.
7- Eghbalsaied, S.h., K. Ghaedi., S. Shahmoradi., A. Pirestani., T. saeidi., H. Amini., L. Nicol., and A. McNielly. 2012. Presence of SNPs in GDF9 mRNA of Iranian Afshari sheep. International Journal fertilization Sterility. 5: 225-230.
8- Hanrahan, J.P., S.M. Gregan., P. Mulsant., M. Mullen., G.H. Davis., R. Powell., and S.M. Galloway. 2004. Mutations in the genes for oocyte derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare sheep (Ovisaries). Biology of Reproduction. 70: 900–909.
9- Juengel, J.L., N.L.Hudson., L. Whiting., and K.P. McNatty. 2004. Effects of immunization against bone morphog-enetic protein 15 and growth different-iation factor 9 on pregnancy in Ewes. Biology of Reproduction. 70: 557-561.
10- Luis, V., P. Ricardo., M.T. Tejedor., L. Adolfo., and S. Isidro. 2009. A 17 bp deletion in the BoneMorphogenetic Protein 15 (BMP15) gene is associated to increased prolificacy in the Rasa Arago-nesa sheep breed. Animal Reproduction Science. 110: 139–146.
11- Mishra, A.K., A.L. Arora, S. Kumar, L.L.Prince. 2009. Studies on effect of Booroola (FecB) genotype on lifetime ewes’ productivity efficiency, litter size and number of weaned lambs in Garole×Malpura sheep. Animal Reprodution Science,113:293–298.
12- Moradband, F., G. Rahimi., and M. Gholizadeh. 2011. Association of polymorphisms in Fecundity Genes of GDF9, BMP15 and BMP15-1B with Litter Size in Iranian Baluchi Sheep.Asian-Australian Journal Animal Science. 9: 1179 – 1183.
13- Msoffe, P.L.M., M.M.A. Mtambo., U.M. Minga., H.R. Juul-Madsen., P.S. Gwakisa. 2005. Genetic structure among the local chicken ecotypes of Tanzania based on microsatellite DNA typing. African Journal of biotechnology. 4: 768-771.
14- Nicol, L., S.C. Bishop., R. Pong-Wong., C.h. Bendixen., L.E. Holm., S.M. Rhind., and A.S. McNeilly. 2009. Homozygosity for a single base-pair mutation in the oocytespecific GDF9 gene results in sterility in Thoka sheep. Science Reproduction Fertilization. 138:921–933.
15- Otsuka, F., Z. Yao., T.H. Lee., S. Yamamoto., G.F. Erickson., and S. Shimasaki. 2000 Bone morphogenetic Protein-15. Identification of target cells and biological functions. Journal Biology Chemistry. 50: 39523–39528.
16- Rahimi, A., M. mohagheghdolatabadi. 2013. Identification of single nucleotide polymorphisms (SNPs) in exon 2 the Growth Differentiation Factor 9 (GDF9) gene in Bahmani and ghashghai sheep. Animal and birds congress, north Iran,1454-1458. (In Persian)
17- Rajaei, M.A. 2005. Study of Genetic diversity for Japan quil population using microsatellite Markers.MSc Thesis of Animal science.Faculty of Agriculture. Tarbiat Modares University.
18- Sadighi, M., K.J. Bodensteiner., A.E. Beattie., and S.M. Galloway. 2002 Genetic mapping of ovine growth differentiation factor 9 (GDF9) to sheep chromosome 5. Animal Genetics. 33: 244–245.
19- Soleimani, B and G.h. Rahimi. 2010.Studyof effect GDF9 gene in litter size and weight quality in sheep sanjabi. Modern Genetics Journal. 5:53-59 (In Persian)
20- Yazdi, M. H., Engstom., G., Nasholm., A. Jonansson., K., Jorjani., H. and Liljedahl. L. E. 1997. Genetic parameters for lamb weight at different age and wool productioninBaluchi sheep. Journal of Animal science. 65(2): 247- 255.
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