Polymorphism In Growth Hormone Gene Sequence From Microminipig (MMP) With Direct Sequencing PCR
Shedage Kishor Ashok, Yasushi Kuwabara, Shunnosuke Abe, Eugene Hayato Morita
Index Terms: Amino acids, Direct DNA sequencing, Growth Hormone, Microminipig, Polymorphism.
Abstract: Pig Growth Hormone regulates growth, development and various metabolic activities in body. It releases from anterior pituatory gland of hypothalamus region of brain. Till now the sequence variation for GH has been carried out only in normal pig and minipigs. This is first report showing GH gene sequence variation in Microminipigs, smallest pig in the world developed by Fuji Nojo Service, Japan which weighs about 6-7 kg at its maturity. In present study, coding and intronic region of GH was determined for polymorphism with direct sequencing PCR. Coding region of pig GH is 651 bp and represents 216 amino acids containing initial 26 amino residues codes for signal peptide. Current investigation showed that, there were total of 6 synonymous and 4 non synonymous changes found, among them, two of each were common in MMP. Non synonymous changes were Val9 and Gln22 were majorly observed in signal peptide of MMP GH. Further analysis study showed that, intronic region was highly polymorphic and sequence variability observed was 7.5 %, 2.7 % and 26.2 % in normal pigs, minipigs and Microminipigs respectively.
 D.L. Roith, C. Bondy, S. Yakar, J.L. Liu, and A. Butler, “The Somatomedin Hypothesis: 2001,” Endocrine Reviews, vol. 22, no. 1, pp. 53–74, 2001.
 M.F. Scanlon, B.G. Issa, and C. Dieguez, “Regulation of growth hormone secretion,” Hormone Research, vol 46, no. 4-5. pp. 149-154, 1996.
 M. Yerle, Y. Mansais, P.D. Thomsen, and J. Gellin, “Localization of the porcine growth hormone gene to chromosome 12p1.2-->p1.5,” Animal Genetics, vol 24, no. 2, pp. 129-131, 1993.
 P.D. Vize, and J.R. Wells, “Isolation and characterization of the porcine growth hormone gene,” Gene, vol. 55, no. 2-3, pp. 339-344, 1987.
 S.S. Abdel-Meguid, H.S. Shieh, W.W. Smith, H.E. Dayringer, B.N. Violand, and L.A. Bentle, “Three dimensional structure of a genetically engineered variant of porcine growth hormone,” Proceddings of the National Academy of Sciences USA, vol. 84, pp. 6434-6437, 1987.
 M.J. Waters, H.N. Hoang, D.P. Fairlie, R.A. Pelekanos, and R.J. Brown, “New insights into growth hormone action,” Journal of Molecular Endocrinology, vol. 36, pp. 1–7, 2006.
 A.J. Brooks, J.W. Wooh, K.A. Tunny, and M.J. Waters, “Growth hormone receptor; mechanism of action,” The International Journal of Biochemistry and Cell Biology, vol. 40, no. 10, pp. 1984-1989, 2008.
 D.A. Faria, S.E.F. Guimaraes, P.S. Lopes, V.P. Aldrin, R.P. Samuel, et al, “Association between G316A growth hormone polymorphism and economic traits in pigs,” Genetics and Molecular Biology, vol. 29, no.4, pp. 634–640, 2006.
 D.W. Yandell, and T.P. Dryja, “Detection of DNA sequence polymorphisms by enzymatic amplification and direct genomic sequencing,” The American Journal of Human Genetics, vol. 45, pp. 547-555, 1989.
 D.G. Wang, J.B. Fan, C.J. Siao, A. Berno, P. Young, and et al., “Large-scale identification, mapping, and genotyping of single-nucleotide polymorphisms in the human genome,” Science, vol. 280, no. 5366, pp. 1077-1082, 1998.
 N. Kaneko, K. Itoh, A. Sugiyama, and Y. Izumi, “Microminipig, a non-rodent experimental animal optimized for life science research: preface. 2011,” Journal of Pharmacological Sciences, vol. 115, no. 2, pp. 112 – 114, 2011.
 R.N. Kirkwood, P.A. Thacker, and B. Laarveld, “The influence of growth hormone injections on the endocrine and metabolic status of gilts,” Domestic Animal Endocrinology, vol. 6, no. 2, pp. 167 – 176, 1989.
 J. Leger, C. Garel, A. Fjellestad-Paulsen, M. Hassan, and P. Czernichow, “Human growth hormone treatment of short stature children born small for gestational age: effect on muscle and adipose tissue mass during a 3-year treatment period and after 1 year's withdrawal,” The Journal of Clinical Endocrinology and Metabolism, vol. 83, no. 10, pp. 3512-3516, 1998.
 D.S. Lough, L.D. Muller, R.S. Kensinger, L.C.Jr Griel, and C.D. Azzara, “Effect of exogenous bovine somatotropin on mammary lipid metabolism and milk yield in lactating dairy cows,” Journal of Dairy Science, vol. 72, no. 6, pp. 1469 – 1476, 1989.
 N. Shimoda, T. Tashiro, H. Yamamori, K. Takagi, N. Nakajima, and I. Ito, “Effects of growth hormone and insulin-like growth factor-1 on protein metabolism, gut morphology, and cell-mediated immunity in burned rats,” Nutrition, vol. 13, no. 6, pp. 540 – 546, 1997.
 J. Skarda, “Effect of bovine growth hormone on growth, organ weights, and tissue composition and adipose tissue metabolism in young castrated male goats,” Livestock Production Science, vol. 55, pp. 215 – 225, 1998.
 K. Sangeeta, R.F. Gagel, and J.C. Gilbert, “Direct sequencing of PCR products in agarose gel slices,” Nucleic Acids Research, vol. 22, no. 16, pp. 3425-3426, 1994.
 C. Missarelli, L. Herrera, V. Mericq, and P. Carvallo, “Two different 5’ splice site mutations in the growth hormone gene causing autosomal dominant growth hormone deficiency,” Human Genetics, vol. 101, no. 1, pp. 113-117, 1997.
 Binder G, Brown M, and Parks JS, Mechanisms responsible for dominant expression of human growth hormone gene mutations,” The Journal of Clinical Endocrinology and Metabolism, vol. 81, no. 11, pp. 4047- 4050, 1996.
 J.D Cogan, B. Ramel, M. Lehto, J 3rd Phillips, M. Prince, R.M. Blizzard, T.J. de Ravel, M. Brammert, and L. Groop, “A recurring dominant negative mutation causes autosomal dominant growth hormone deficiency--a clinical research center study,” The Journal of Clinical Endocrinology and Metabolism, vol. 80, no. 12, pp. 3591- 3595, 1995.
 J.D Cogan, J 3rd Phillips, N. Sakati, H. Frisch, E. Schober, R.D. Milner, Heterogeneous growth hormone (GH) gene mutations in familial GH deficiency,” The Journal of Clinical Endocrinology and Metabolism,” vol. 76, no. 5, pp. 1224 – 1228, 1993.
 P. Duquesnoy, S. Amselem, M. Gourmelen, Y. Le Bouc, and M. Goossens, “A frameshift mutation causing isolated growth hormone deficiency type IA (Abstract),” American Journal of Medicine , vol. 47,pp. 110, 1990.
 J.T Li, Y.L. Mu, L. Zhang, S.L. Yang, K. Li, and S.T. Feng, “New mutations in growth hormone and receptor genes from Chinese Wuzhishan miniature pig,” Acta Agriculturae Scandinavica Section A Animal Science, vol. 57, no. 2, pp. 97-100, 2007.
 J.Z. Deng, L.L. Hao, M.T. Li, S. Lang, Y.Z. Zeng, S.C. Liu, and Y.L. Zhang, “Growth hormone and receptor gene mutations in Chinese Banna Miniature pig,” Animal Cells and systems, 15, no. 4, pp. 310- 314, 2011.
 L.I. Jing, R. Xuq-Qin, and J.F. Wang, “Identification and function of the growth hormone gene in Rongjiang pig of China,” Acta Physiologica Sinica, vol. 58, no. 3, pp. 217-224, 2006.
 D.J. O’mahony, H. Wang, D.J. McConnell, F. JIA, L. XIA, and S. QI, “Polymorphism in porcine somatotropin cDNA sequences,” Animal Genetics, vol. 20, pp. 313-316, 1989.
 Y. Hasegawa, K. Fujii, M. Yamada, Y. Igarashi, K. Tachibana, T. Tanaka, K. Onigata, Y. Nishi, S. Kato, T. Hasegawa, “Identification of Novel Human GH-1 Gene Polymorphisms that are Associated with Growth Hormone Secretion and Height,” The Journal of Clinical Endocrinology and Metabolism, vol. 85, no. 3, pp. 1290-1295, 2000.
 D.S. Millar DS, M. Horan, N.A. Chuzhanova, and D.N. Cooper, “Characterisation of a functional intronic polymorphism in the human growth hormone (GH1) gene,” Human Genomics, vol. 4, no. 5 pp. 289–301, 2010.
 N. J. Larsen and V. H. Nielsen, “Characterisation of a functional intronic polymorphism in the human growth hormone (GH1) gene,” Human Genomics, vol. 8, no. 2 pp. 151–166, 2010.