Research Article
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investigation of FecXI mutation by PCR-RFLP method in Awassi sheep breed

Year 2022, Volume: 11 Issue: 2, 88 - 93, 16.12.2022

Oğuz Ağyar Koray Kırıkçı

Abstract

The identification of mutations affecting multiple births in sheep is critical for the efficacy of genetic breeding studies. Before carrying out breeding studies with marker-assisted selection (MAS), it is necessary to identify major or potential novel mutations and determine their effects at the level of the breed. The aim of this study was to investigate the FecXI (Inverdale) mutation in the Awassi sheep breed using the PCR-RFLP technique. The study used blood samples from 50 ewes collected from a pure breeding farm in the Adiyaman province of Turkey. To isolate genomic DNA from blood, a commercial isolation kit was utilized. The 154 base pair area of the BMP15 gene where the mutation was located was amplified by PCR, and the FecX mutation was analyzed in all PCR products using the PCR-RFLP technique. The FecXI mutation was not found in the Awassi sheep tested using PCR-RFLP. It is recommended that the major and recently found novel mutations with multiple births be investigated by different molecular methods in the Awassi sheep breed.

Keywords

sheep multiple births Awassi PCR-RFLP FecXI BMP15

Supporting Institution

-

Project Number

-

Thanks

The authors thank Mr. Oktay YAVAS, who is a trainer of molecular biology and genetics at Kırşehir Ahi Evran University, for his help in laboratory studies.

References

  • Abdoli, R., Mirhoseini, S. Z., Ghavi Hossein‐Zadeh, N., Zamani, P., & Gondro, C. (2018). Genome‐wide association study to identify genomic regions affecting prolificacy in Lori‐Bakhtiari sheep. Animal Genetics, 49(5), 488-491.
  • Abdullah, A.Y., Husein, M.Q., Kridli, R.T.(2002).Protocols for estrus synchronization in Awassi ewes under arid conditions, Asian Austral. J. Anim. Sci., 15, pp. 957-962, 10.1111/rda.12733.
  • Al-Nafie, A. T., Al-Thuwaini, T. M., & Al-Shuhaib, M. B. S. (2022). A novel association between hemoglobin subunit beta gene and reproductive performance in Awassi ewes. Journal of the Saudi Society of Agricultural Sciences, 21(1), 1-7.
  • Çelikeloğlu, K., Tekerli, M., Erdoğan, M., Koçak, S., Hacan, Ö., & Bozkurt, Z. (2021). An investigation of the effects of BMPR1B, BMP15, and GDF9 genes on litter size in Ramlıç and Dağlıç sheep. Archives Animal Breeding, 64(1), 223-230.
  • Davis, G.H., Balakrishnan, L., Ross, I.K., Wilson, T., Galloway, S.M., Lumsden, B.M., Hanrahan, J.P., Mullen, M., Mao, X.Z., Wang, G.L. (2006). Investigation of the Booroola (FecB) and Inverdale (FecXI) mutations in 21 prolific breeds and strains of sheep sampled in 13 countries. Anim Reprod Sci, 92 (1-2): 87-96.dosage-sensitive manner. Nat Genet, 25(3): 279-283.
  • Davis, G.H., McEwan, J.C., Fennessy, P.F., Dodds, K.G., Farquhar, P.A. (1991). Evidence for the presence of a major gene influencing ovulation rate on the X-chromosome of sheep. Biol Reprod 1991; 44:620–624. Davis, G. H. (2005). Major genes affecting ovulation rate in sheep. Genetics Selection Evolution, 37(Suppl. 1), S11-S23
  • Galal, S., Gursoy, O. and Shaat., I. (2008). Awassi sheep as a genetic resource and efforts for their genetic improvement—a review, Small Ruminant Research, 79, 99–108.
  • Galloway, S.M., McNatty, K.P., Cambridge, L.M., Laitinen, M.P., Juengel, J.L., Jokiranta, T.S., McLaren, R.J., Luiro, K., Dodds, K.G., Montgomery, G.W. (2000). Mutations in an oocyte-derived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner. Nature genetics, 25(3), 279–283. https://doi.org/10.1038/77033.
  • Gedik, Y. (2021). Screening for Inverdale (FecXI) Mutation in BMP15 Gene in Prolific Turkish Awassi Sheep. Black Sea Journal of Agriculture, 4(4), 130-132.
  • Gootwine, E., Reicher, S., & Rozov, A. (2008). Prolificacy and lamb survival at birth in Awassi and Assaf sheep carrying the FecB (Booroola) mutation. Animal reproduction science, 108(3-4), 402-411.
  • Gürsel, E. (2011). Determination of BMP-15, BMPR-1B and GDF-9 gene mutations of the indigenous sheep breeds in Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 17(5), 725-729.
  • Gürsoy, O. (2005). Small ruminant breeds of Turkey. In Characterization of Small Ruminant Breeds in West Asia , North Africa, Vol 1: West Asia, Luis Iñiguez, Ed, ICARDA, Aleppo, Syria, 239-416.
  • Karslı, T., Balcıoğlu, M. S. (2010). Türkiye'de yetiştirilen altı yerli koyun ırkında BMPR-IB (Booroola) Geninde FecB allel varlığının PCR-RFLP yöntemiyle araştırılması. Kafkas Universitesi Veteriner Fakultesi Dergisi, 17(5), 1033-1036.
  • Karsli, T., Sahin, E., Karsli, B. A., Alkan, S., Balcioglu, M. S. (2012). An investigation of mutations (FecXG, FecXI, FecXH, FecXB) on BMP-15 gene in some local sheep breeds raised in Turkey. Akdeniz Üniv. Ziraat Fak, 25(1), 29-33.
  • Kirikci, K., & Çam, M. (2020). Türkiye yöresel yeni koyun tipi Of koyunlarında GDF9 (FecG1) gen polimorfizmin PCR-RFLP yöntemi ile Araştırılması. Manas Journal of Agriculture Veterinary and Life Sciences, 10(2), 98-102.
  • Kirikci, K. (2022). Polymorphism of the Calpastatin (CAST) and Growth Differentiation Factor 9 (GDF9) genes in Akkaraman Sheep Breed. Hayvansal Üretim, 63(1), 21-26.
  • Kirikci, K., Cam, M.A., & Mercan, L. (2021). Investigation of G1 (c. 260G> A) polymorphism in exon 1 of GDF9 gene in Turkish sheep breed Karayaka, Turkish Journal of Veterinary & Animal Sciences, 45(1), 191-197.
  • Mahdavi, M., Nanekarani, S., & Hosseini, S. D. (2014). Mutation in BMPR-IB gene is associated with litter size in Iranian Kalehkoohi sheep. Animal reproduction science, 147(3-4), 93-98.
  • McNatty, K. P., Juengel, J. L., Wilson, T., Galloway, S. M., Davis, G. H., Hudson, N. L., Moeller, C. L., Cranfield, M., Reader, K. L., Laitinen, M. P., Groome, N. P., Sawyer, H. R., & Ritvos, O. (2003). Oocyte-derived growth factors and ovulation rate in sheep. Reproduction (Cambridge, England). Supplement, 61, 339–351.
  • Mohammed, M. M., Al-Thuwaini, T. M., & Al-Shuhaib, M. B. S. (2022). A novel p. K116Q SNP in the OLR1 gene and its relation to fecundity in Awassi ewes. Theriogenology, 184, 185-190.
  • Nagdy, H., Mahmoud, K. G. M., Kandiel, M. M., Helmy, N. A., Ibrahim, S. S., Nawito, M. F., & Othman, O. E. (2018). PCR-RFLP of bone morphogenetic protein 15 (BMP15/FecX) gene as a candidate for prolificacy in sheep." International journal of veterinary science and medicine, 6(sup1), S68-S72.
  • Notter, D.R. (2008). Genetic Aspects of Reproduction in Sheep. Reprod. Domest. Anim. 43, 22-128.
  • Rothschild, M., Jacobson, C., Vaske, D., Tuggle, C., Wang, L., Short, T., Eckardt, G., Sasaki, S., Vincent, A., McLaren, D., Southwood, O., van der Steen, H., Mileham, A., & Plastow, G. (1996). The estrogen receptor locus is associated with a major gene influencing litter size in pigs. Proceedings of the National Academy of Sciences of the United States of America, 93(1), 201–205. https://doi.org/10.1073/pnas.93.1.201.
  • Rummel, T., Valle Zárate, A. and Gootwine, E. (2005). The world wide gene flow of the Improved Awassi and Assaf sheep breeds from Israel. In: Gene flow in animal genetic resources. A study on status, impact and trends (Institute of Animal Production in the Tropics and Subtropics of the University of Hohenheim, Germany).
  • Thieme, O., Karazeybek, M., Özbayat, H., İbrahim, H., Sözmen, R. (1999).Performance of village sheep flocks in Central AnatoliaII. Fertility and productivity of ewes. Turkish Journal of Veterinary and Animal Sciences. 23 (2); 175-181. https://journals.tubitak.gov.tr/veterinary/vol23/iss2/13. Üstüner, H., Oğan, M. M. (2013). Main productive performance of Awassi sheep in the Central Anatolian region of Turkey. Turkish Journal of Veterinary and Animal Sciences. 37; 271-276, 10.3906/vet-1205-13.

İvesi koyun ırkında FecXI mutasyonunun PCR-RFLP yöntemi ile araştırılması

Year 2022, Volume: 11 Issue: 2, 88 - 93, 16.12.2022

Oğuz Ağyar Koray Kırıkçı

Abstract

Koyunlarda çoklu doğumu etkileyen mutasyonların tanımlanması genetik ıslah çalışmalarının etkinliği açısından oldukça önemlidir. Markör destekli seleksiyon (MAS) ile ıslah çalışmalarının yapılabilmesi için öncelikli olarak majör veya olası yeni mutasyonların tanımlanması ve ırk düzeyinde etkisinin ortaya konması gerekmektedir. Bu çalışmada İvesi koyun ırkında FecXI (Inverdale) mutasyonunun PCR-RFLP yöntemi ile araştırılması amaçlanmıştır. Çalışmanın materyalini, Adıyaman ilinde saf yetiştiriciliği yapılan bir işletmeden alınan 50 baş İvesi koyun ırkına ait kan örnekleri oluşturmuştur. Kandan genomik DNA’nın elde edilmesinde ticari izolasyon kiti kullanılmıştır. Mutasyonun bulunduğu BMP15 geninin 154 baz çiftlik bölgesi PCR ile çoğaltılmış ve sonrasında tüm PCR ürünlerinde FecXI mutasyonu PCR-RFLP yöntemi ile araştırılmıştır. PCR-RFLP sonuçları, incelenen İvesi koyunlarının FecXI mutasyonunu taşımadığını göstermiştir. İvesi koyun ırkında çoklu doğum ile ilişkili majör ve tanımlanan yeni mutasyonların farklı moleküler yöntemler ile araştırılması önerilmektedir.

Keywords

Koyun Çoklu doğum İvesi PCR-RFLP FecXI BMP15

Project Number

-

References

  • Abdoli, R., Mirhoseini, S. Z., Ghavi Hossein‐Zadeh, N., Zamani, P., & Gondro, C. (2018). Genome‐wide association study to identify genomic regions affecting prolificacy in Lori‐Bakhtiari sheep. Animal Genetics, 49(5), 488-491.
  • Abdullah, A.Y., Husein, M.Q., Kridli, R.T.(2002).Protocols for estrus synchronization in Awassi ewes under arid conditions, Asian Austral. J. Anim. Sci., 15, pp. 957-962, 10.1111/rda.12733.
  • Al-Nafie, A. T., Al-Thuwaini, T. M., & Al-Shuhaib, M. B. S. (2022). A novel association between hemoglobin subunit beta gene and reproductive performance in Awassi ewes. Journal of the Saudi Society of Agricultural Sciences, 21(1), 1-7.
  • Çelikeloğlu, K., Tekerli, M., Erdoğan, M., Koçak, S., Hacan, Ö., & Bozkurt, Z. (2021). An investigation of the effects of BMPR1B, BMP15, and GDF9 genes on litter size in Ramlıç and Dağlıç sheep. Archives Animal Breeding, 64(1), 223-230.
  • Davis, G.H., Balakrishnan, L., Ross, I.K., Wilson, T., Galloway, S.M., Lumsden, B.M., Hanrahan, J.P., Mullen, M., Mao, X.Z., Wang, G.L. (2006). Investigation of the Booroola (FecB) and Inverdale (FecXI) mutations in 21 prolific breeds and strains of sheep sampled in 13 countries. Anim Reprod Sci, 92 (1-2): 87-96.dosage-sensitive manner. Nat Genet, 25(3): 279-283.
  • Davis, G.H., McEwan, J.C., Fennessy, P.F., Dodds, K.G., Farquhar, P.A. (1991). Evidence for the presence of a major gene influencing ovulation rate on the X-chromosome of sheep. Biol Reprod 1991; 44:620–624. Davis, G. H. (2005). Major genes affecting ovulation rate in sheep. Genetics Selection Evolution, 37(Suppl. 1), S11-S23
  • Galal, S., Gursoy, O. and Shaat., I. (2008). Awassi sheep as a genetic resource and efforts for their genetic improvement—a review, Small Ruminant Research, 79, 99–108.
  • Galloway, S.M., McNatty, K.P., Cambridge, L.M., Laitinen, M.P., Juengel, J.L., Jokiranta, T.S., McLaren, R.J., Luiro, K., Dodds, K.G., Montgomery, G.W. (2000). Mutations in an oocyte-derived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner. Nature genetics, 25(3), 279–283. https://doi.org/10.1038/77033.
  • Gedik, Y. (2021). Screening for Inverdale (FecXI) Mutation in BMP15 Gene in Prolific Turkish Awassi Sheep. Black Sea Journal of Agriculture, 4(4), 130-132.
  • Gootwine, E., Reicher, S., & Rozov, A. (2008). Prolificacy and lamb survival at birth in Awassi and Assaf sheep carrying the FecB (Booroola) mutation. Animal reproduction science, 108(3-4), 402-411.
  • Gürsel, E. (2011). Determination of BMP-15, BMPR-1B and GDF-9 gene mutations of the indigenous sheep breeds in Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 17(5), 725-729.
  • Gürsoy, O. (2005). Small ruminant breeds of Turkey. In Characterization of Small Ruminant Breeds in West Asia , North Africa, Vol 1: West Asia, Luis Iñiguez, Ed, ICARDA, Aleppo, Syria, 239-416.
  • Karslı, T., Balcıoğlu, M. S. (2010). Türkiye'de yetiştirilen altı yerli koyun ırkında BMPR-IB (Booroola) Geninde FecB allel varlığının PCR-RFLP yöntemiyle araştırılması. Kafkas Universitesi Veteriner Fakultesi Dergisi, 17(5), 1033-1036.
  • Karsli, T., Sahin, E., Karsli, B. A., Alkan, S., Balcioglu, M. S. (2012). An investigation of mutations (FecXG, FecXI, FecXH, FecXB) on BMP-15 gene in some local sheep breeds raised in Turkey. Akdeniz Üniv. Ziraat Fak, 25(1), 29-33.
  • Kirikci, K., & Çam, M. (2020). Türkiye yöresel yeni koyun tipi Of koyunlarında GDF9 (FecG1) gen polimorfizmin PCR-RFLP yöntemi ile Araştırılması. Manas Journal of Agriculture Veterinary and Life Sciences, 10(2), 98-102.
  • Kirikci, K. (2022). Polymorphism of the Calpastatin (CAST) and Growth Differentiation Factor 9 (GDF9) genes in Akkaraman Sheep Breed. Hayvansal Üretim, 63(1), 21-26.
  • Kirikci, K., Cam, M.A., & Mercan, L. (2021). Investigation of G1 (c. 260G> A) polymorphism in exon 1 of GDF9 gene in Turkish sheep breed Karayaka, Turkish Journal of Veterinary & Animal Sciences, 45(1), 191-197.
  • Mahdavi, M., Nanekarani, S., & Hosseini, S. D. (2014). Mutation in BMPR-IB gene is associated with litter size in Iranian Kalehkoohi sheep. Animal reproduction science, 147(3-4), 93-98.
  • McNatty, K. P., Juengel, J. L., Wilson, T., Galloway, S. M., Davis, G. H., Hudson, N. L., Moeller, C. L., Cranfield, M., Reader, K. L., Laitinen, M. P., Groome, N. P., Sawyer, H. R., & Ritvos, O. (2003). Oocyte-derived growth factors and ovulation rate in sheep. Reproduction (Cambridge, England). Supplement, 61, 339–351.
  • Mohammed, M. M., Al-Thuwaini, T. M., & Al-Shuhaib, M. B. S. (2022). A novel p. K116Q SNP in the OLR1 gene and its relation to fecundity in Awassi ewes. Theriogenology, 184, 185-190.
  • Nagdy, H., Mahmoud, K. G. M., Kandiel, M. M., Helmy, N. A., Ibrahim, S. S., Nawito, M. F., & Othman, O. E. (2018). PCR-RFLP of bone morphogenetic protein 15 (BMP15/FecX) gene as a candidate for prolificacy in sheep." International journal of veterinary science and medicine, 6(sup1), S68-S72.
  • Notter, D.R. (2008). Genetic Aspects of Reproduction in Sheep. Reprod. Domest. Anim. 43, 22-128.
  • Rothschild, M., Jacobson, C., Vaske, D., Tuggle, C., Wang, L., Short, T., Eckardt, G., Sasaki, S., Vincent, A., McLaren, D., Southwood, O., van der Steen, H., Mileham, A., & Plastow, G. (1996). The estrogen receptor locus is associated with a major gene influencing litter size in pigs. Proceedings of the National Academy of Sciences of the United States of America, 93(1), 201–205. https://doi.org/10.1073/pnas.93.1.201.
  • Rummel, T., Valle Zárate, A. and Gootwine, E. (2005). The world wide gene flow of the Improved Awassi and Assaf sheep breeds from Israel. In: Gene flow in animal genetic resources. A study on status, impact and trends (Institute of Animal Production in the Tropics and Subtropics of the University of Hohenheim, Germany).
  • Thieme, O., Karazeybek, M., Özbayat, H., İbrahim, H., Sözmen, R. (1999).Performance of village sheep flocks in Central AnatoliaII. Fertility and productivity of ewes. Turkish Journal of Veterinary and Animal Sciences. 23 (2); 175-181. https://journals.tubitak.gov.tr/veterinary/vol23/iss2/13. Üstüner, H., Oğan, M. M. (2013). Main productive performance of Awassi sheep in the Central Anatolian region of Turkey. Turkish Journal of Veterinary and Animal Sciences. 37; 271-276, 10.3906/vet-1205-13.
There are 25 citations in total.

Details

Primary Language English
Subjects Zootechny (Other)
Journal Section Research Article
Authors

Oğuz Ağyar

Koray Kırıkçı

Project Number -
Publication Date December 16, 2022
Published in Issue Year 2022 Volume: 11 Issue: 2

Cite

APA Ağyar, O., & Kırıkçı, K. (2022). investigation of FecXI mutation by PCR-RFLP method in Awassi sheep breed. Bahri Dağdaş Hayvancılık Araştırma Dergisi, 11(2), 88-93.
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