Gökkuşağı Alabalığı (Oncorhynchus mykiss, Walbaum 1792) Antalya Kültür Popülasyonu Büyüme Hormonu I Geni İzolasyonu ve Karakterizasyonu

Year 2020, Issue: 20, 223 - 232, 31.12.2020

Mesut Yılmaz , Mehmet Özbaş , Mehmet Akif Kılıç

https://doi.org/10.31590/ejosat.781631

Abstract

Bu çalışmada, Antalya İli’nde yetiştirilen gökkuşağı alabalığı kültür popülasyonundan büyüme hormonu I geni izole ve karakterize edilmiştir. Bu amaçla kültür popülasyonundan 10 birey örnek olarak alınmıştır. Örneklenen balıkların kas dokularından toplam DNA izole edilmiştir. Polimeraz zincir reaksiyonunda kullanılmak üzere primerler dizayn edilmiş ve hedef gen bölgesi çoğaltılmıştır. İzole edilen genin dizi analizi yapılmış ve 4591 nükleotitten oluştuğu tespit edilmiştir. Referans diziye kıyasla 2 transisyonel, 14 transversiyonel ve 38 insersiyonel baza karşılık gelen 54 nükleotitten oluşan mutasyon bulunmuştur. Tüm nükleotit farklılıkları genin intron bölgelerinde tespit edilmiştir. Genin intron bölgelerinde yer alan putatif hormon yanıt elemanları (HRE) değerlendirildiğinde ise daha önce raporlanan dizi ile tamamen uyumlu olduğu görülmüştür. Elde edilen veriler filogenetik ve gen aktarım çalışmalarında kullanılabilecek niteliktedir.

Keywords

Büyüme hormonu geni, rtghI, HRE, Gökkuşağı alabalığı, Oncorhynchus mykiss

Supporting Institution

Akdeniz Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi

Project Number

2009.03.0121.012

Thanks

Bu çalışmada uygulanan prosedürler Akdeniz Üniversitesi Tıp Fakültesi Hayvan Bakım-Kullanım ve Hayvan Deneyleri Etik Kurulu tarafından değerlendirilmiş ve 2009.09.40 nolu karar ile etik olarak uygunluğu onaylanmıştır. Ayrıca çalışma Akdeniz Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından 2009.03.0121.012 no’lu proje ile desteklenen MY’a ait doktora tezinden üretilmiştir.

Isolation and Characterization of Growth Hormone I Gene from Antalya Culture Population of Rainbow Trout (Oncorhynchus mykiss, Walbaum 1792)

Abstract

In this study, the growth hormone I gene was isolated and characterized from the rainbow trout culture population reared in Antalya Province. For this purpose, 10 individuals from the culture population were sampled. Total DNA was isolated from the muscle tissue of fish. Primers used in the polymerase chain reaction were designed and the target gene region was amplified. Sequence analysis of the isolated gene was performed and it was determined that the gene consisted of 4591 nucleotides. Compared to the reference sequence, 54 nucleotide differences were found corresponding to 2 transitional, 14 transversional and 38 insertional bases. All nucleotide variations are located in the intron regions of the gene. When the putative hormone response elements (HRE) were evaluated, they were seen to be completely compatible with the previously reported sequence. The data obtained in this study can be used in phylogenetic and gene transfer studies.

Keywords

Growth hormone gene, rtghI, HRE, Rainbow trout, Oncorhynchus mykiss

Project Number

2009.03.0121.012

References

• Agellon, L.B. & Chen, T.T. (1986). Rainbow trout growth. hormone: molecular cloning of cDNA and expression in Escherichia coli. DNA, 5(6), 463-471.
• Agellon, L.B., Davies, S.L., Chen, T.T. & Powers, D.A. (1988). Structure of a fish (Rainbow trout) growth hormone gene and ıts evolutionary ımplications. Proceedings of the National Academy of Sciences of the United States of America, 85, 5136-5140.
• Barlow, J. W., Voz, M. L., Eliard, P. H., Mathy-Harter, M., De Nayer, P., Economidis, I. V., Martial, J. A. & Rousseau, G. G. (1986). Thyroid hormone receptors bind to defined regions of the growth hormone and placental lactogen genes. Proceedings of the National Academy of Sciences, 83(23), 9021–9025.
• Ber, R., & Daniel, V. (1993). Sequence analysis suggests a recent duplication of the growth hormone-enco ding gene in Tilapia nilotica. Gene 125, 143-150.
• Björnsson, B.T. (1997). The biology of salmon growth hormone: from daylight to dominance. Fish Physiology and Biochemistry,17: 9–24.
• Brody, J.R. & Kern, S.E. (2004). History and principles of conductive media for standard DNA electrophoresis. Analytical Biochemistry, 333(1), 1-13.
• Carvalho, G. R. (1993). Evolutionary aspects of fish distribution: genetic variability and adaptation. Journal of Fish Biology, 43(sa), 53–73.
• Dieffenbach, C.W., Lowe, T.M.J. & Dveksler, G.S. (1995). General concepts for PCR primer design ın: PCR Primer, A Laboratory Manual, Dieffenbach CW, Dveksler GS Ed., Cold Spring Harbor Laboratory Press, New York.
• Filiz, E. & Koç, İ. (2011). Bitki Biyoteknolojisinde Moleküler Markörler. GOÜ, Ziraat Fakültesi Dergisi, 28(2), 207-214.
• Freeland, J. R. (2008). Ekologia Molekularna. Wydawnictwo Naukowe PWN, Warszawa.
• Gerstein, M.B., Bruce, C., Rozowsky, J.S., Zheng, D., Du, J., Korbel, J.O., Emanuelsson, O., Zhang, Z.D., Weissman, S. & Snyder, M. (2007). What is a gene, post-ENCODE? History and updated definition. Genome Research. 17 (6), 669-681.
• Hall, T.A. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98. Hüttenhofer, A., Schattner, P. & Polacek, N. (2005). Non-coding RNAs: hope or hype?. Trends Genet. 21 (5), 289–97.
• Johansen, B., Johnsen, O.C., &Valla, S. (1989). The complete nucleotide sequence of the growth hormone gene from Atlantic salmon (Salmo salar). Gene 77, 317-324.
• Kalendar, R., Lee, D. & Schulman, A.H. (2009). Fast PCR Software for PCR Primer and Probe Design and Repeat Search. Genes, Genomes and Genomics, 3(1), 1-14.
• Kaplan, S. L. (1999) Hormonal regulation of growth and metabolic effects of growth hormone. In: Kostyo JL, Goodman HM (Eds) Handbook of Physiology. The endocrine system, Vol V: Hormonal control of growth. Oxford University Press, New York.
• Kennedy, S. & Oswald, N. (2011). PCR Troubleshooting and Optimization: The Essential Guide. Caister Academic Press, Norfolk, UK.
• Klug, W.S., Cummings, M.R., Palladino, M. A. & Spencer, C. A. (2018). Genetik Kavramlar (11. Baskı). Palme Yayıncılık, Ankara.
• Kocher, T.D. & Carleton, K.L. (1997). Base Substitution in Fish Mitochondrial DNA: Patterns and Rates: içinde Molecular Systematics of Fishes, pp 13-24. Academic Press, California.
• Male, R., Nerlaand, A.H., Lorens, J.B., Telle, W., Lossius, I., & Totland, G.K. (1992). The complete nucleotide sequence of the Atlantic salmon growth hormone I gene. Biochim. Biophys. Acta 1130, 345-348.
• Marins, L. F., Levy, J. A., Folch, J. M. & Sanchez, A. (2003). A growth hormone-based phylogenetic analysis of euteleostean fishes including a representative species of the Atheriniformes Order, Odontesthes argentinensis. Genetics and Molecular Biology, 26 (3), 295-300.
• Mori, T., Deguchi, F. & Ueno, K. (2001). Differential Expression of GH1 and GH2 Genes by Competitive RT-PCR in Rainbow Trout Pituitary. General and Comprative Endocrinology, 123, 137–143.
• Mount, S.M. (1982). A catalogue of splice junction sequences. Nucleic Acids Res. 10, 459-472.
• Nelson, D. L. & Cox, M. M. (2017). Lehninger Principles of Biochemistry (7th Ed.). W.H. Freeman and Company, New York.
• Nicoll, C. S., Rodgers, B. D. & Kelly, K. M. (1999) Hormonal regulation of growth and development of nonmammalian vertebrates. In: Kotsyo JL, Goodman HM (Eds) Handbook of physiology. The endocrine system, Vol V, Hormonal control of growth. Oxford University Press, NY.
• Rajesh, R. & Majumdar, K. C. (2007). A comparative account of the structure of the growth hormone encoding gene and genetic interrelationship in six species of the genus Labeo. Fish Physiol Biochem, 33, 311–333.
• Rothan, H. A., Ser Huy, T., & Mohamed, Z. (2014). Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone inPichia pastoris. The Scientific World Journal, 1–6.
• Sekine, S., Mizukami, T., Nishi, T., Kuwana, Y., Saito, A., Sato, M., Itoh, S. & Kawauchi, H. (1985). Cloning and expression of cDNA for salmon growth hormone in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America, 82, 4306–4310.
• Slater, E. P., Rabenau, O., Karin, M., Baxter, J. D., & Beato, M. (1985). Glucocorticoid receptor binding and activation of a heterologous promoter by dexamethasone by the first intron of the human growth hormone gene. Molecular and Cellular Biology, 5(11), 2984–2992.
• Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22(22), 4673–4680.
• Wagner, G.F., Fargher, R.C., Brown, J.C. & Mckeown, B.A. (1985). Further chacterization of growth hormone from chum salmon (Oncorhynchus keta). General and Comparative Endocrinology, 60, 27-34.
• Xia, J., Xia, K. & Jiang, S. (2008). Complete mitochondrial DNA sequence of the yellowfin seabream Acanthopagrus latus and a genomic comparison among closely related sparid species. Mitochondrial DNA 19(4), 385-393.
• YANG, B., CHAN, K., LIN C. & CHEN, T.T. (1997). Characterization of rainbow trout (Oncorhynchus mykiss) growth hormone 1 gene and the promoter region of growth hormone 2 gene. Archives of Biochemistry Biophysic, 340, 359–368.
• Yılmaz, M. 2012. Dağ Alası (Salmo trutta macrostıgma Dumeril, 1858) için β-aktin Promotör-rtghı İfade Vektörünün Hazırlanması. Doktora Tezi, Akdeniz Üniversitesi, Fen Bilimleri Enstitüsü, Antalya.
• Yowe, D.L., & Epping, R.J. (1995). Cloning of the barramundi growth hormone-encod ing gene: a comparative analysis of higher and lower vertebrate GH genes. Gene 162, 255-259.