Türkiye'de Yetiştirilen Pekin Ördeklerinde Büyüme Hormonu Geni Polimorfizmi

Yıl 2022, , 798 - 806, 23.12.2022

Candan Eriş Cengiz Elmacı

https://doi.org/10.33462/jotaf.1089941

Öz

Moleküler genetik alanındaki gelişmeler çiftlik hayvanlarının morfolojik ve fizyolojik birçok özelliklerini etkileyen genlerin veya belirteçlerin tanımlanmasına yol açmıştır. Bu özelliklerin moleküler temeli, fonksiyonel genomik yöntemlerle ortaya çıkarılmakta ve belirteç destekli seleksiyon (MAS) yoluyla çiftlik hayvanlarında genetik ıslah programlarını geliştirme fırsatları sağlamaktadır. Hayvanlarda çeşitli verim ve üreme performanslarının MAS yoluyla arttırılması amacıyla çeşitli aday genler üzerinde durulmaktadır. Üzerinde durulan bu aday genlerden birisi de büyüme hormonu genidir. Bu çalışmanın temel amacı, Pekin ördeklerinde büyüme hormonu geninin 2., 3. ve 4. intronlarındaki polimorfizmleri araştırmaktır. Bu amaçla Bursa Uludağ Üniversitesi Ziraat Fakültesi Tarımsal Uygulama ve Araştırma Çiftliğinde yetiştirilen her iki cinsiyetten (79 erkek ve 38 dişi) toplam 117 adet yetişkin Pekin ördeğinden alınan kan örnekleri materyal olarak kullanılmıştır. Büyüme hormonu geninin 2., 3. ve 4. intronlarındaki genetik polimorfizm, üç primer çifti ile PCR-RFLP yönteminde BsmF1 restriksiyon enzimi kullanılarak araştırılmıştır. Kesim sonrası elde edilen DNA parçalarının ayrımı için %2’lik agaroz jel elektroforezi uygulanmıştır. PCR-RFLP analizleri sonucunda sadece 2. intron bölgesinde polimorfizm belirlenmiştir. 2. intron bölgesinden çoğaltılan parçanın BsmF1 restriksiyon enzimi ile kesimi sonucu bu bölgede TT (765 bç), CT (765, 593 ve 172 bç) ve CC (593 ve 172 bç) olmak üzere üç genotip tespit edilmiş ve T ve C allellerinin frekansları da sırasıyla 0,75 ve 0,25 olarak hesaplanmıştır. Çalışılan populasyonda büyüme hormonu geni 2. intron bölgesinde gözlenen değerlerle beklenen değerler arasındaki farklılık istatistiksel olarak önemli olmadığından (P>0.05) ele alınan populasyonun Hardy-Weinberg dengesinde olduğu anlaşılmıştır. 3. ve 4. intronlarda ise genetik varyasyon gözlenmemiştir. Büyüme hormonu genin 3. ve 4. intronlarına ait sırasıyla 442 bç ve 1378 bç’lik bölgelere ait PCR ürünleri de BsmF1 restriksiyon enzimi ile muamele edilmiş, ancak bu enzime ait kesim bölgesine rastlanmamış ve bu nedenle üzerinde çalışılan populasyonda 3.ve 4.intronda tek tip genotip elde edilmiştir. Dolayısıyla bu iki bölgenin BsmF1 lokusu bakımından monomorfik olduğu anlaşılmıştır. Bu ve önceki çalışmalarda büyüme hormonu geninin 2. intron bölgesinde gösterilen genotipik varyasyon, bu bölge ile çeşitli verimler arasındaki ilişkileri konu alan daha çok çalışma yapılması gerekliliğini de ortaya koymaktadır.

Anahtar Kelimeler

Büyüme hormonu (BH) geni, PCR, RFLP, Genetik varyasyon, Polimorfizm, Pekin ördeği, Polymorphism

Growth Hormone Gene Polymorphism in Pekin Ducks Reared in Turkey

Öz

Advances in molecular genetics have led to the identification of genes or markers that affect many morphological and physiological characteristics of farm animals. The molecular basis of these traits is revealed by functional genomic methods and provides opportunities to develop genetic improvement programs in farm animals through marker assisted selection (MAS). Several candidate genes are being studied with the aim of increasing various yield and reproductive performances in animals through MAS. One of these candidate genes is the growth hormone gene. The main purpose of this study was to investigate polymorphisms in the 2nd, 3rd and 4th introns of the growth hormone gene in Pekin ducks. For this purpose, blood samples taken from a total of 117 adult Pekin ducks of both sexes (79 males and 38 females) raised in Bursa Uludag University Faculty of Agriculture Agricultural Application and Research Farm were used as material. Genetic polymorphism in the 2nd, 3rd and 4th introns of the growth hormone gene was investigated using the BsmF1 restriction enzyme in the PCR-RFLP method with three primer pairs. Samples were carried out in 2% agarose gel electrophoresis to separate the DNA fragments obtained after the digestion. As a result of the researches, polymorphism was determined only in the fragment amplified from the 2nd intron region. As a result of the digestion of the amplified fragment from the 2nd intron region with BsmF1 restriction enzyme, three genotypes, namely TT (765 bp), CT (765, 593 and 172 bp) and CC (593 and 172 bp) were determined in this region and the frequencies of the T and C alleles were calculated as 0.75 and 0.25, respectively. Since the difference between the values observed in the 2nd intron region of the growth hormone gene and the expected values in the studied population was not statistically significant (P>0.05), it was understood that the population in studied was in Hardy-Weinberg equilibrium. No genetic variation was observed in the 3rd and 4th introns. PCR products belonging to the 442 bp and 1378 bp regions of the 3rd and 4th introns of the growth hormone gene, respectively, were also treated with BsmF1 restriction enzyme, but the cleavage region of this enzyme was not found, so there was only one genotype was obtained in the 3rd and 4th introns of the studied population. Therefore, it was understood that these two regions are monomorphic in terms of BsmF1 locus. The genotypic variation shown in the 2nd intron region of the growth hormone gene in this and previous studies also reveals the need for further studies on the relationships between this region and various yields.

Anahtar Kelimeler

Growth hormone (GH) gene, PCR, RFLP, Genetic variation, Pekin duck, Polimorphism

Kaynakça

• Anonim, (2018). Tarım ve Orman Bakanlığı, TAGEM, Ar-Ge İnovasyon, Kanatlı hayvancılık sektör politika belgesi 2018-2022. https://www.tarimorman.gov.tr/TAGEM/Belgeler/yayin/Kanatl%C4%B1%20Hayvanc%C4%B1l%C4%B1k%20Sekt%C3%B6r%20Politika%20Belgesi%202018-2022.pdf (Erişim tarihi:31/05/2022)
• Barta, A., Richards, R.I., Baxter, J.D., Shine, J. (1981). Primary structure and evolution of rat growth hormone gene. Proceedings of the National Academy of Sciences, 78(8) 4867-4871.
• Bauman, D.E. (1999). Bovine somatotropin and lactation: from basic science to commercial application. Domestic Animal Endocrinology, 17(2-3): 101-116.
• Bhattacharya, T.K., Chatterjee, R.N. (2013). Polymorphism of the myostatin gene and its association with growth traits in chicken. Poultry Science, 92(4): 910-915.
• Byrne, C.R., Wilson, B.W., Ward, K.A. (1987). The isolation and characterization of the ovine growth hormone gene. Australian Journal of Biological Sciences, 40(4): 459-468.
• Chang, M., Cheng, Y., Huang, M. (2012). The SNP genotypes of growth hormone gene associated with reproductive traits in Tsaiya Ducks. Reproduction in Domestic Animals, 47(4): 568–573.
• Das, P., Meyer, L., Seyfert, H.M., Brockmann, G., Schwerin, M. (1996). Structure of the growth hormone-encoding gene and its promoter in mice. Gene, 169(2): 209-213.
• Dettori, M.L. Pazzola, M., Pira, E., Paschino, P., Vacca, G.M. (2015). The sheep growth hormone gene polymorphism and its effects on milk traits. Journal of Dairy Research, 82(2):169-176.
• Du, S.J., Devlin, R.H., Hew, C.L. (1993). Genomic structure of growth hormone genes in chinook salmon (Oncorhynchus tshawytscha): presence of two functional genes, GH-I and GH-II, and a male-specific pseudogene, GH-psi. DNA and Cell Biology, 12(8): 739-751.
• Gala, R.R. (1991). Prolactin and growth hormone in the regulation of the immune system. Experimental Biology and Medicine, 198(1):513-527.
• Ge, X., Yu, J., Jiang, H. (2012). Growth hormone stimulates protein synthesis in bovine skeletal muscles cells without altering insulin-like growth factor-I mRNA expression. Journal of Animal Sciences, 90(4): 1126-1133.
• Ghelghachi, A.A., Reza, S.H., Lak, A. (2013). Association of growth hormone gene polymorphism with growth and fatness traits in Arian broilers. International Journal of Biosciences, 3(12): 216-220.
• Hiyama, G., Okabayashi, H., Kansaku, N., Tanaka, K. (2012). Genetic variation in the growth hormone promoter region of Anas platyrhynchos, a duck native to Myanmar. Journal of Poultry Sciences, 49(4): 245-248.
• Huang, Y.Z., He, H., Zhang, Z.Y., Sun, Y.J., Li, M.X., Lan, X.Y., Leic, Z., Zhang, C.L., Chen, H. (2013). Relationship of polymorphisms within ZBED6 gene and growth traits in beef cattle. Gene, 526(2): 107-111.
• Hull, K.L., Harvey, S. (2001). Growth hormone: roles in female reproduction. Journal of Endocrinology, 168(1): 1-23. Jafari, A., Pakdel, A., Esmailkhanian, S. (2015). Growth hormone gene polymorphism in two Iranian Native Fowls (Short Communication). Poultry Science Journal, 3 (1): 99-104.
• Jing, L.I., Ran, X.-Q., Wang, J.F. (2006). Identification and function of the growth hormone gene in Rongjiang pig of China. Acta Physiologica Sinica, 58(3): 217-224.
• Johari, S., Setiati, N., Sidadolog, J.H.P., Hartatik, T., Yuwanta, T. (2013). The gene effect of growth hormone on body weight and egg production in divergent selection for five generation of Japanese Quail (Coturnix coturnix japonica). International Journal of Poultry Science, 12(8): 489-494.
• Kansaku, N., Nakada, A., Okabayashi, H., Guemene, D., Kuhnlein, U., Zadworny, D., Shimada, K. (2003). DNA polymorphism in the chicken growth hormone gene: Association with egg production. Animal Science Journal., 74(3):243-244.
• Kansaku, N., Soma, A., Furukawa, S., Hiyama, G., Okabayashii, H., Guemene, D., Kühnlein, U., Zadworny, D. (2008). Sequence of the domestic duck (Anas platyrhynchos) growth hormone-encoding gene and genetic variation in the promoter region. Animal Science Journal, 79(2): 163-170.
• Kuhnlein, U., Ni, L., Weigend, S., Gavora, J.S., Fairfull, W., Zadworny, D. (1997). DNA polymorphisms in the chicken growth hormone gene: Response to selection for disease resistance and association with egg production. Animal Genetics, 28(2): 116-123.
• Lucy, M.C., Hauser, S.D., Eppard, P.J., Krivi, G.G., Clark, J.H., Bauman, D.E., Collier, R.J. (1993). Variants of somatotropin in cattle: gene frequencies in major dairy breeds and associated milk production. Domestic Animal Endocrinology, 10(4):325-333.
• Malveiro, E., Pereira, M., Marques, P.X., Santos, I.C., Belo, C., Renaville, R., Cravador, A. (2001). Polymorphisms at the five exons of the growth hormone gene in the algarvia goat: possible association with milk traits. Small Ruminant Research, 41(2): 163-170.
• Mazurowski, A., Frieske, A., Kokoszynski, D., Mroczkowski, S., Bernacki, Z., Wilkanowska, A. (2015). Examination of growth hormone (GH) gene polymorphism and its association with body weight and selected body dimensions in ducks. Folia Biologica (Kraków), 63(1): 43-50.
• Nie, Q., Ip, S.C.Y., Zhang, X., Leung, F.C., Yang, G. (2002). New variations in intron 4 of growth hormone gene in Chinese Native Chickens. Journal of Heredity, 93(4): 277-279.
• Nie, Q., Sun, B., Zhang, D., Luo, C., Ishag, N.A., Lei, M., Yang, G., Zhang, X. (2005). High diversity of the chicken growth hormone gene and effects on growth and carcass traits. Journal of Heredity, 96(6):698–703.
• Ohlsson, C., Bengtsson, B.A., Isaksson, O.G., Adreassen, T.T., Slootweg, M.C. (1998). Growth hormone and bone. Endocrine Reviews, 19(1): 55-79.
• Sheng-Wai, X., Wen-Bin, B., Jun, H., Jun-Hua, C., Jing-Ting, S., Gua-Hong, C. (2007). Polymorphic analysis of intron 2 and 3 of growth hormone gene in duck. Hereditas (Beijing), 29(4): 438-442.
• Soysal, M.İ., Kök, S., Gürcan, E.K. (2005). An Investigation on the distribution in erythrocytes potassium polymorphisms in buffaloes. Journal of Tekirdağ Agricultural Faculty. 2(2):189-193.
• Stephen, C.Y., Zhang, X., Leung, F. (2001). Genomic growth hormone gene polymorphisms in native Chinese chickens. Experimental Biology and Medicine, 226(5):458-462.
• Supakorn, C., Pralomkarn, W. (2013). Genetic polymorphisms of growth hormone (GH), insulin-like growth factor 1 (IGF-1) and diacylglycerol acyltransferase 2 (DGAT-2) genes and their effect on birth weight and weaning weight in goats. Philippine Agricultural Scientist, 96(1): 18-25.
• Şekeroğlu, A., Özen, N. (1997). Tarım ve Köyişleri Bakanlığı üretme istasyonlarında kanatlı hayvan yetiştiriciliği. Akdeniz Üniversitesi Ziraat Fakültesi Dergisi, 10:336-344.
• Tanaka, M., Hosokowa, Y., Watahıkı, M., Nakashıma, K. (1992). Structure of the chicken growth hormone-encoding gene and its promoter region. Gene, 112(2): 235-239.
• Wang, C., Liang Z., Yu, W., Feng, Y., Peng, X., Gong, Y., Li, S. (2011). Polymorphism of the prolactin gene and its association with egg productiontraits in native Chinese ducks. South African Journal of Animal Science, 41(1): 63-69.
• Woychik, R.P., Camper, S.A., Lyons, R.H., Horowitz, S., Goodwin, E.C., Rottman, F.M. (1982). Cloning and nucleotide sequencing of the bovine growth hormone gene. Nucleic Acids Research, 10(22): 7197-7210.
• Wu, Y., Pan, A.L., Pi, J.S., Pu, Y.J., Du, J.P., Liang, Z.H., Shen, J. (2012). One novel SNP of growth hormone gene and its associations with growth and carcass traits in ducks. Molecular Biology Reports, 39(8):8027–8033.
• Wu, X., Yan , M.J., Lıan , S.Y., Liu, X.T., Li, A. (2014). GH gene polymorphisms and expression associated with egg laying in Muscovy ducks (Cairina moschata). Hereditas, 151(1): 14–19.
• Yeh, F., Yang, R.C., Boyle, T. (2000). Popgene (v.1,32), Microsoft Windows-based freeware for Population Genetic analysis. http://www.ualberta.ca/*fyeh/Pop32.exe.
• Zhang, X.L., Jiang, X., Liu, Y.P., Du, H.R., Zhu, Q. (2007). Identification of AvaI polymorphisms in the third intron of GH gene and their associations with abdominal fat in chickens. Poultry Science, 86(6): 1079–1083.
• Zhang, Y., Zhu, Z., Xu, Q., Chen, G. (2014). Association of polymorphisms of exon 2 of the growth hormone gene with production performance in Huoyan goose. International Journal of Molecular Sciences, 15(1): 670-683.
• Zhang, D.X., Xu, Z.Q., He, J., Ji, C.L., Zhang, Y., Zhang, X.Q. (2015). Polymorphisms in the 5’-flanking regions of the GH, PRL and Pit-1 genes with Muscovy duck egg production. Journal of Animal Science, 93(1): 28-34.
• Zhao, W.M., Zhao, R.X., Qiao, N., Xu, Q., Huang, Z.Y., Li, X., Zhang, Y., Chen, G.H. (2011). Association of GH polymorphisms with growth traits in goose. Journal of Animal and Veterinary Advances, 10(6): 692-697.