請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8698完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳金洌 | |
| dc.contributor.author | Chiou-Yueh Lee | en |
| dc.contributor.author | 李秋月 | zh_TW |
| dc.date.accessioned | 2021-05-20T19:59:53Z | - |
| dc.date.available | 2014-03-10 | |
| dc.date.available | 2021-05-20T19:59:53Z | - |
| dc.date.copyright | 2010-03-10 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-02-11 | |
| dc.identifier.citation | 參 考 文 獻
Acosta, J., Carpio, Y., Borroto, I., Gonzalez. O. and Estrada, M.P. 2005. Myostatin gene silenced by RNAi show a zebrafish giant phenotype. J. Biotechnol. 119:324-331. Akiyoshi, S., Inoue, H., Hanai, J., Kusanagi, K., Nemoto, N., Miyazono, K. and Kawabata, M. 1999. c-Ski acts as a transcriptional co-repressor in transforming growth factor-beta signaling through interaction with smads. J Biol Chem. 274:35269-35277. Amali, A.A., Lin, C.J.F., Chen, Y.H., Wang, W.L., Gong, H.Y., Lee, C.Y., Ko, Y.L., Lu, J.K., Her, G.M., Chen, T.T. and Wu, J.L. 2004. Up-regulation of muscle-specific transcription factors during embryonic somitogenesis of zebrafish (Danio rerio) by knock-down of myostatin-1. Dev. Dyn. 229:847-856. Amali, A.A., Lin, C.J.F., Chen, Y.H., Wang, W.L., Gong, H.Y., Rekha, R.D., Lu, J.K., Chen, T.T. and Wu, J.L. 2008. Overexpression of myostatin2 in zebrafish reduces the expression of dystrophin associated protein complex (DAPC) chich leads to muscle dystrophy. J. Biomed. Sci. 15:595-604. Anderson, S.B., Goldberg, A.L. and Whitman, M. 2008. Identification of a novel pool of extracellular pro-myostatin in skeletal muscle. J. Biol. Chem. 283:7027-7035. Artaza, J.N., Reisz-Porszasz, S., Dow, J.S., Kloner, R.A., Tsao, J., Bhasin, S. and Gonzalez-Cadavid, N.F. 2007. Alterations in myostatin expression are associated with changes in cardiac left ventricular mass but not ejection fraction in the mouse. Journal of Endocrinology 194:63-76. Bass, J., Oldham, J., Sharma, M., and Kambadur, R. 1999. Growth factors controlling muscle development. Domest. Anim. Endocrinol. 17:191-197. Benezra, R., Davis, R.L., Lockshon, D., Turner, D.L., and Weintraub, H. 1990. The protein Id: a negative regulator of helix-loop-helix DNA-binding proteins. Cell 61:49-59. Biga, P.R., Roberts, S.B., Iliev, D.B., McCauley, L.A.R., Moon, J.S., Collodi, P., and Goetz, F.W. 2005. The isolation, characterization, and expression of a novel GDF11 gene and a second myostatin form in zebrafish, Danio rerio. Comp. Biochem. Physiol. 141:218-230. Blackwell, T.K. and Weintraub, H. 1990. Differences and similarities in DNA-binding preferences of MyoD and E2A protein complexes revealed by binding site selection. Science. 250(4984):1104-1110. Braun, T., Rudnicki, M.A., Arnold, H.-H., and Jaenish, R. 1992. Targeted inactivation of the muscle regulatory gene Myf-5 results in abnormal rib development and perinatal death. Cell 71:369-382. Camp, E. and Lardelli, M. 2001. Tyrosinase gene expression in zebrafish embryos. Dev Genes Evol. 211:150-153. Casas, E., Keele, J.W., Fahrenkrug, S.C., Smith, T.P., Cundiff, L.V., and Stone, R.T. 1999. Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele. J. Anim. Sci. 77:1686-1692. Corrtnik, D., Et al., 1992. The retinoblastoma protein and the regulation of cell cyling. Trends. Biochem. Sci. 17:312-315. Coustry, F., Maity, S.N. and de Crombrugghe, B. 1995. Studies on transcription activation by the multimeric CCAAT-binding factor CBF. J. Biol. Chem. 270:468-475. Crisa, A., Marchitelli, C., Savarese, M.C. and Valentini, A. 2003. Sequence analysis of myostatin promoter in cattle. Cytogenet. Genome Res. 102:48-52. Davis R.L., Cheng, P.F., Lassar, A.B. and Weintraub, H. 1990. The myoD DNA binding domain contains a recognition code for muscle-specific gene activation. Cell 60:733-746. Davis, R.L., Weintraub, H., and Lassar, A.B. 1987. Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51:987-1000. De Santis, C., Evans, B.S., Smith-Keune, C. and Jerry, D.R. 2008. Molecular characterization, tissue expression and sequence variability of the barramundi (Lates calcarifer) myostatin gene. BMC Genomics 9:82. Fahrenkrug, S.C., Casas, E., Keele, J.W., and Smith, T.P. 1999. Technical note: direct genotyping of the double-muscling locus(mh) in Piedmontese and Belgian Blue cattle by fluorescent PCR. J. Anim. Sci. 77:2028-2030. Ferrell, R.E., Conte, V., Lawrence, E.C., Roth, S.M., Hagberg, J.M., and Hurley, B.F. 1999. Frequent sequence variation in the human myostatin (GDF8) gene as a marker for analytsis of muscle-related phenotypes. Genomics 62:203-207. Funkenstein, B. and Rebhan, Y. 2007. Expression, purification, renaturation and activation of fish myostatin expressed in Escherichia coli: Facilitation of refolding and activity inhibition by myostatin prodomain. Protein Expr Purif. 54: 54-65. Gaussin V, Depre C. 2005. Myostatin, the cardiac chalone of insulin-like growth factor-1. Cardiovascular Research 68:347-349. Gonzalez-Cadavid, N.F., Taylor, W.E., Yarasheski, K., Sinha-Hikim, I., Ma, K., Ezzat, S., Shen, R., Lalani, R., Asa, S., Mamita, M., Nair, G., Arver, S., and Bhasin, S. 1998. Organization of the human myostatin gene and expression in healthy men and HIV-infected men with muscle wasting. Proc. Natl. Acad. Sci. 95:14938-14943. Grobet, L., Martin, L.J., Poncelet, D., Pirottin, D., Brouwers, B., Riquet, J., Schoeberlein, A., Dunner, S., Menissier, F., Massabanda, J., Fries, R., Hanset, R., and Georges, M. 1997. A deletion in the bovine myostatin gene causes the double-muscled phenotype in cattle. Nat. Genet. 17:71-74. Hasty, P., Bradley, A., Morris, J.H., Edmondson, D.G., Venuti, J.M., Olson, E., and Klein, W.H. 1993. Muscle deficiency and neonatal death in mice with a targeted mutation in the myogenin gene. Nature 364:501-506. Hiebert, S.W., Chellappan, S.P., Horowitz, J.M. and Nevins, J.R. 1992. The interaction of RB with E2F coincides with an inhibition of the transcriptional activity of E2F. Genes Dev. 6:177-185. Hill, J.J., Davies, M.V., Pearson, A.A., Wang, J.H., Hewick, R.M., Wolfman, N.M. and Qiu, Y. 2002. The myostatin propeptide and the follistatin-related gene are inhibitory binding proteins of myostatin in normal serum. J. Biol. Chem. 277: 40735-40741. Hollingsworth, R.E., J.R., Hensey C.E. and Lee, W.H. 1993. Retinoblastoma protein and the cell cycle. Curr. Opin. Genet. Dev. 3:55-62. Jeanplong, F., Bass, J.J., Smith, H.K., Kirk, S.P., Kambadur, R., Sharma, M., Oldham, J.M. 2003. Prolonged underfeeding of sheep increases myostatin and myogenic regulatory factor Myf-5 in skeletal muscle while IGF-I and myogenin are repressed. J. Endocrinol. 176:425-437. Jen, Y., Weintraub, H., and Benezra, R. 1992. Overexpression of Id protein inhibits the muscle differentiation program; In vivo association of Id and E2A proteins. Genes. Dev. 6:1466-1479. Ji, S., Losinski, R.L., Cornelius, S.G., Frank, G.R., Willis, G.M., Gerrard, D.E., Depreux, F.F., and Spurlock, M.E. 1998. Myostatin expression in procine tissues: tissue specificity and developmental and postnatal regulation. Am. J. Physiol. 275:R1265-R1273. Jones, N. 1990. Transcriptional regulation by dimerization: Two sides to an incestuous relationship. Cell 61:9-11. Kambadur, R., Sharma, M., Smith, T.P., and Bass, J. J. 1997. Mutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle. Genome. Res. 7:910-916. Kerr, T., Roalson, E.H. and Rodgers, B.D. 2005. Phylogenetic analysis of the myostatin gene sub-family and the differential expression of a novel member in zebrafish. Evol. Dev. 7:390-400. Kobayashi, N., Goto, K., Horiguchi, K., Nagata, M., Kawata, M., Miyazawa, K., Saitoh, M. and Miyazono, K. 2007. c-Ski activates MyoD in the nucleus of myoblastic cells through suppression of histone deacetylases. Genes Cells. 12:375-385. Koli, K., Saharinen, J., Hyytiäinen, M., Penttinen, C. and Keski-Oja, J. 2001. Latency, activation, and binding proteins of TGF-beta. Microsc. Res. Tech. 52:354-62. Kollias, H.D., Perry, R.L., Miyake, T., Aziz, A. and McDermott, J.C. 2006. Smad7 promotes and enhances skeletal muscle differentiation. Mol. Cell Biol. 26:6248-6260. Langley, B., Thomas, M., Bishop, A., Sharma, M., Gilmour, S. and Kambadur, R. 2002. Myostatin inhibits myoblast differentiation by down regulating myoD expression. J. Biol. Chem. 27751:49831-49840. Lassar, A.B., Buskin, J.N., Lockshon, D., Davis, R.L., Apone, S., Hauschka, S.D., and Weintraub, H. 1989. MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer. Cell 58:823-831. Lassar, A.B., Paterson, B.M., and Weintraub, H. 1986. Transfection of a DNA locus that mediated the conversion of C3H10T1/2 fibroblasts into myoblasts. Cell 47:649-656. Lawlor, M.A. and Rotwein, P. 2000. Insulin-like growth factor-mediated muscle cell survival central roles for Akt and cyclin-dependent kinase inhibitor p21. Mol. Cell Biol. 20:8983-8995. Lee, S.J. 2004. Regulation of muscle mass by myostatin. Annu. Rev. Cell Biol. 20:61-86. Lee, S.J. and McPherron, A.C. 2001. Regulation of myostatin activity and myuscle growth. Proc. Natl. Acac. Sci. 98:9306-9311. Lyons, G.E., and Buckingham, M.E. 1993. Developmental regulation of myogenesis in the mouse. Semin. Dev. Biol. 3:243-253. Ma, K., Mallidis, C., Artaza, J., Taylor, W., Gonzalez-Cadavid, N. and Bhasin, S. 2001. Characterization of 5’-regulatory region of human myostatin gene: regulation by dexamethasone in vitro. Am. J. Physiol. Endocrinol. Metab. 281:E1128-E1136. Maccatrozzo, L., Bargelloni, L., Patarnello, P., Radelli, G., Mascarello, F. and Patarnello, T. 2002. Characterization of the myostatin gene and a linked microsatellite marker in shi drum (Umbrina cirrosa, Sciaenidae). Aquaculture 205:49-60. Massagué, J. 1990. The transforming growth factor-beta family. Annu. Rev. Cell Biol. 6:597-641. Mckoy, G., Bicknel,l K.A, Patel, K, Brooks G. 2007. Developmental expression of myostatin in cardiomyocytes and its effect on foetal and neonatal rat cardiomyocyte proliferation. Cardiovascular Research 74: 304-312. McPherron, A.C., and Lee, D.J. 1997. Double muscleing in cattle due to mutations in the myostatin gene. Proc. Natl. Acad. Sci. 94:12457-12461. McPherron, A.C., Lawler, A.M., and Lee, S.J. 1997. Regulation of skeletal muscle mass in mice by a new TGF- superfamily member. Nature 387:83-90. McPherron, A.C. and Lee, D.J. 2002. Suppression of body fat acumulation in myostatin-deficient mice. J. Clin. Invest. 109:595-601. Modesti, P.A., Vanni, s., Bertolozzi, I., Cecioni, I., Lumachi, C., Perna, A.M., Boddi, M. and Gensini, G.F. 2004. Different growth factor activation in the right and left ventricles in experimental volume overload. Hypertension. 43:101-108 Morissette, M.R., Cook, S.A., Foo, S., McKo,y G., Ashida, N., Novikov, M., Scherrer-Crosbie, M., Li, L., Matsui, T., Brooks, G. and Rosenzweig, A. 2006. Myostatin regulates cardiomyocyte growth through modulation of Akt signaling. Circulation Research 99:15-24. Nabeshima, Y., Hanaoka, K., Hayasaka, M., Esumi, E., Li, S., Nonaka, I., and Nabeshima, Y. 1993. Myogenin gene disruption results in perinatal lethality because of severe muscle defect. Nature 364:532-535. Naya, F.J., Wu, C., Richardson, J.A., Overbeek, P. and Olson, E.N. 1999. Transcriptional activity of MEF2 during mouse embryogenesis monitored with a MEF2-dependent transgene. Development. 126:2045-2052. Nigg, E.A. 1995. Cyclin-dependent protein kinases: key regulators of the eukaryotic cell cycle. Bioessays 17:471-480. Nishimura T, Oyama K, Kishioka Y, Wakamatsu J, Hattori A. 2007. Spatiotemporal expression of decorin and myostatin during rat skeletal muscle development. Biochem. Biophys. Re.s Commun. 361: 896-902. Olson, E.N. 1990. MyoD family: a paradigm for development. Genes Dev. 4:1454-1461. Østbye, T.K. Galloway, T.F., Nielsen, C., Gabestad, I., Bardal, T. and Andersen, O. 2001. The two myostatin genes of Atlantic salmon (Salmo salar) are expressed in a variety of tissues. Eur. J. Biochem. 268:5249-5257. Pie, M.R and Alvares, L.E. 2006. Evolution of myostatin in vertebrates: is there evidence for positive selection. Mol. Phylogenet. Evol. 41:730-734. Pirottin, D., Poncelet, D., Grobet, L., Royo, L.J., Brouwers, B., Masabanda, J., Takeda, H., Fries, R., Sugimoto, Y., Womack, J.E., Junner, S., and Georges, M. 1999. High-resolution, human-bovine comparative mapping based on a colsed YAC contig spanning the bovine mh locus. Mamm. Genome. 10:289-293. Rios, R., Carneiro, I., Arce, V.M., and Devesa, J. 2001. Myostatin regulates cell survival during C2C12 myogenesis. Biochem. Biophys. Res. Commun. 280:561-566. Roberts, S.B., and Goetz, F.W. 2001. Differential skeletal muscle expression of myostatin across teleost species, and the isolation of multiple myostatin isoforms. FEBS. Lett. 491:212-216. Rodgers, B.D. and Weber, G..M. 2001. Sequence conservation among fish myostatin orthologues and the characterization of two additional cDNA colnes form Morone saxatilis and Morone Americana. Comp. Biochem. Physiol. Part B 129:597-603. Rodgers, B.D., Weber, G.M., Sullivan, C.V., and Levine, M.A. 2001. Isolation and haracterization of myostatin complementary deoxyribonucleic acid clones form two commerically important fish: Oreochromis mossambicus and Morone chrysops. Endocrinology 142:1412-418. Rudnicki, M.A., Braun, T., Hinuma, S., and Jaenisch, R. 1992. Inactivation of MyoD in mice leads to up-regulation of the myogenic HLHgene Myf-5 and results in apparently normal muscle development. Cell 71:383-390. Rudnicki, M.A., Schnegelsberg, P.N.J., Stead, r.H., Braun, T., Arnold, H.-H., and Jaenisch, R. 1993. Myo D or Myf-5 is required for the formation of skeletal muscle. Cell 75:1351-1359. Salerno, M.S., Thomas, M., Forbes, D., Watson, T. Kambadur, R. and Sharma, M. 2004. Molecular analysis of fiber-type specific expression of murine myostatin promoter. Am. J. Physiol. Cell Physiol. 287:C1031-C1040. Schuelke, M., Wagner, K.R., Stolz, L.E., Hubner, C., Riebel, T., Komen, W., Braun, T., Tobin, J.F. and Lee, S.J. 2004. Myostatin mutation associated with gross muscle hypertrophy in a child. N. Engl. J. Med. 350: 2682-2688. Sharma, M., Kambadur, R., Matthews, K.G., Somers, W.G., Devlin, G.P., Conaglen, J.V., Fowke, P.J. and Bass, J.J. 1999. Myostatin, a transformaing growth factor-beta superfamily member, is expressed in heart muscle and is upregulated in cardiomyocytes after infarct. J. Cell. Physiol. 180:1-9. Smith, T.P., Lopez-Corrales, N.L., Kappes, S.M., and Sonstegard, t. S. 1997. Myostatin maps to the interval containing the bovine mh locus. Mamm.Genome. 8:742-744. Sonstegard, t.S., Kappes, s.M., Keele, J.W., and smith, T.P. 1998. Refinement of bovine chromosome 2 linkage map near the mh locus reveals rearrangements between the bovine and human genomes. Anim. Genet. 29:341-347. Spiller, M.P., Kambadur, R., Jeanplong, F., Thomas, M., Martyn, J.K., Bass, J.J. and Sharma, M. 2002. The myostatin gene is a downstream target gene of basic helix-loop-helix transcription factor myoD. Mol. Cell. Biol. 22:7066-7082. Suzuki, H., Yagi, K., Kondo, M., Kato, M., Miyazono, K. and Miyazawa, K. 2004. c-Ski inhibits the TGF-beta signaling pathway through stabilization of inactive Smad complexes on Smad-binding elements. Oncogene. 23:5068-5076. Taylor, S.M., and Jones P.A. 1979. Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5azacytidine. Cell 17:771-779. Taylor, W.E., Bhasin, S., Artaza, J., Byhower, F., Azam, M., Willard, D.H., Jr, Kull, F.C., and Jr, Gonzalez-Cadavid, N. 2001. Myostatin inhibits cell proliferation and protein synthesis in C2C12 muscle cells. Am. J. Physiol. Endocrinol. Metab. 280:E221-E228. Thies, R.S., Chen, T., Davies, M.V., Tomkinson, K.N., Pearson, A.A., Shakey, Q.A. and Wolfman, N.M.. 2001. GDF-8 propeptide binds to GDF-8 and antagonizes biological activity by inhibiting GDF-8 receptor binding. Growth Factors. 18:251-259. Thomas, M., Langley, B., Berry, C., Sharma, M., Kirk, S., Bass, J., and Kambadur, R. 2000. Myostatin, a negative regulator of muscle growth, functions by inhibiting myoblast proliferation. J. Biol. Chem. 275:40235-40243. Venuti, J.M., Morris, J.H., Vivian, J.L., Olson, E.N., and Klein, W. H. 1995. Myogenesis is required for late but not early aspects of myogenesis during muscle development. J. Cell. Biol. 128:563-576. Velleman, S.G,. Liu, X., Eggen, K.H. and Nestor, K.E. 1999. Developmental regulation of proteoglycan synthesis and decorin expression during turkey embryonic skeletal muscle formation. Poult Sci. 78:1619-1626. Wagner, K.R., Liu, X., Chang, X. and Allen, R.E. 2005. Muscle regeneration in the prolonged absence of myostatin. Proc. Natl. Acad. Sci. 102:2519-2524. Wehling, M., Cai, B., and Tidball, J.G.H. 2000. Modulation of myostatin expression during modified muscle use. FASEB. J. 14:103-110. Weintraub, D.J., Prater, C.A. and Dean, D.C. 1992. Retinoblastoma protein switches the E2F site form a positive to negative element. Nature 358:259-261. Weintraub, H., et al., 1991. The MyoD gene family: nodal point during specification of the muscle cell lineage. Science 251:761-766. Weintraub, H., Tapscott, S.J., Davis, R.L., Thayer, M.J., Adam, M.A., Lasser, A.B., and Miller, D. 1989. Activation of muscle-specific genes in pigment, nerve, fat, liver, and fibroblast cell line by forced expression of MyoD. Proc. Natl. Acac. Sci. 86:5434-5438. Westhusin, M. 1997. From mighty mice to mighty cows. Nat. Genet. 17:4-5. Williams, B.A. and Ordahl, C.P. 1994. Pax-3expression in segmental mesoderm marks early stages in myogenic cell specification. Development 120:785-796. Wolfman, N.M., McPherron, A.C., Pappano, W.N., Davies, M.V., Song, K., Tomkinson, K.N., Wright, J.F., Zhao, L., Sebald, S.M., Greenspan, D.S. and Lee, S.J. 2003. Activation of latent myostatin by the BMP-1/tolloid family of metalloproteinases. Proc. Natl. Acad. Sci. 100:15842-15846. Xu, W., Angelis, K., Danielpour, D., Haddad, M.M., Bischof, O., Campisi, J., Stavnezer, E. and Medrano, E.E. 2000. Ski acts as a co-repressor with Smad2 and Smad3 to regulate the response to type beta transforming growth factor. Proc. Natl. Acad. Sci. 97:5924-5929. Xu, C., jWu, G., Zohar, Y. and Du, S.J. 2003. Analysis of myostatin gene structure, expression and function in zebrafish. J. Exp. Biol. 260:4067-4079. Yang, W., Chen, Y., Zhang, Y., Wang, X., Yang, N. and Zhu, D. 2006. Extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase pathway is involved in myostatin-regulated differentiation repression. Cancer Res. 66:1320-1326. Yang, W., Zhang, Y., Li, Y.F., Wu, Z.G. and Zhu, D.H. 2007. Myostatin induces cyclin D1 degradation to cause cell cycle arrest through a phosphatidylinositol 3-kinase/AKT/GSK-3 beta pathway and is antagonized by insulin-like growth factor 1. J. Biol. Chem. 282:3799-3808. Zhu, X., Hadhazy, M., Wehling, M., Tidball, J.G. and McNally, E.M. 2000. Dominant negative myostatin produces hypertrophy without hyperplasia in muscle. FEBS. Lett: 474:71-75. Zhu, X., Topouzis, S., Liang, L.F. and Stotish, R.L. 2004. Myostatin signaling through Smad2, Smad3 and Smad4 is regulated by the inhibitory Smad7 by a negative feedback mechanism. Cytokine 26: 262-272. Zimmers, T.A., Davies, M.V., Koniaris, L.G., Haynes, P., Esquela, A.F., Tomkinson, K.N., McPherron, A.C., Wolfman, N.M. and Lee, S.J. 2002. Induction of cachexia in mice by systemically administered myostatin. Science 296:1486-1488. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8698 | - |
| dc.description.abstract | 哺乳類動物肌肉倍增基因(Myostatin) 缺乏會造成骨骼肌增加及變大導致成 ”肌肉倍增” 表現型，Myostatin 在牛及人類主要表現在骨骼肌中，其突變或缺乏會造成骨骼肌細胞質量及數目的增加，是高等脊椎動物骨骼肌發育與生長之主要負調控因子；對魚類而言，myostatin可在多種組織中表現，如腦、鰓、心臟、卵巢、眼睛、腎臟等器官，故其並非只單純調控肌肉組織生長，可能同時參與多種組織之調控機制。本實驗則分別從斑馬魚基因體基因庫及 24 小時cDNA 基因庫中篩選基因體 DNA 及 cDNA ，分別探討 myostatin 基因之調控與功能。經選殖後得到7 個有訊號的基因體 DNA ，其中有三個選殖株為 myostatin I 基因組，分別為Zg 3-1的13.5Kb 、Zg 15-1的 24Kb 及 Zg 17-3的15Kb，其中以 15-1 包含有 myostatin I基因的啟動子及基因組在內，為了分析啟動子的表現，利用 PCR 增幅方式取得啟動子片段5.9Kb、5.18Kb、3.4Kb、2.85Kb、2.55Kb、1.69Kb 及 0.6Kb 等7 個片段，並將其構築至 pGL3-basic 冷光表現載體及pEGFP-1綠色螢光蛋白載體上，並將 pGL3-basic 冷光表現選殖株轉殖到 C2C12 、ZFL、NIH3T3 細胞中看其表現， 在 C2C12 及 ZFL 細胞中以 3.4Kb 長的啟動子有最大活性， NIH3T3 細胞表現情形則以 0.6 Kb 有最佳活性，將含 pEGFP-MSYN 綠色螢光蛋白選殖株利用顯微注射打入斑馬魚胚胎中，觀察 myostatin 啟動子在生物體之表現情形，結果並無預期之綠色螢光之表現。
另為瞭解myostatin之功能，由斑馬魚 24 小時胚胎所合成的 cDNA 庫所選殖出 myostatin II 並將 exon III 刪除 281 bp利用vector base-RNA方式將其構築至pEGFP-C1 載體，利用顯微注射打入斑馬魚胚胎中，取得一個遺傳穩定myostatin 基因knockdown 的肌肉倍增斑馬魚。結果在胚胎早期發育時為全身發亮，至72 小時開始往肝臟集中至 96 小時則表現於肝臟及脊椎且穩定綠色螢光表現於成魚中，經進一步交配後其子代仍可穩定且持續的表現。利用定量核酸聚合酶連鎖反應分析確認 RNA降低為野生組的30%，肌肉蛋白質含量降為野生組的36.76%；肌肉生成機制中相關標示因子 MyoD、Myogenin、 Mrf4 和 Myf5 之RNA增加為原來的2倍以上；分析第六代的成長速率，發現在4個月大時， 其野生組和轉基因組兩組間平均體重0.38 ± 0.05 公克及0.55 ± 0.11 公克， 轉基因組體重增加了45%；經 t-test 統計分析兩組間彼此有顯著差異， 進一步將一個月的稚魚切片及經 HE 染色分析肌肉纖維細胞大小由野生組平均值230.69 ± 139.04 μm2 增加至轉基因組之428.97 ± 207.2 μm2 平均增加了 85.95％，且兩者間具有顯著差異(p<0.05)。在本實驗中針對斑馬魚 myostatin 進行基因沉默 (gene silence) 可得到穩定遺傳世代表現之轉殖基因魚，這是首次成功得到具有 'double muscle' 表現型特徵的模式斑馬魚。 | zh_TW |
| dc.description.abstract | Myostatin belongs to the transforming growth factor (TGF)-β superfamily, and is a secreted growth and differentiation factor and also a potent negative regulator of skeletal muscle development and growth. The myostatin (MSTN) null phenotype in mammals were characterized by extreme gains in skeletal muscles mass or “double muscle” as the cytokine negatively regulates skeletal muscle growth and development. Several aspects of MSTN biology in the fish were quit different from that in mammals. MSTN homologs had been identified in many different fish species. This diverse patter of expression in fish suggests that the biological actions of MSTN may not be restricted to skeletal muscle but may additionally influence other fish tissues as well. In this study, we had establish the zebrafish genomic library and obtained seven lambda clones by using the zebrafish MSTN I cDNA as probe. After subcloning, analysing and comparison sequence with the documented MSTN I cDNA, we found that Zg3-1 clone included 12.5Kb promoter region and exon I, Zg15-1 including 11.5 Kb promoter, exon I, intron I, exon II, intron II, exon III, intron III and 3’ untranslation region. To analyze the promoter activity of myostatin I gene, we ligated 7 different DNA fragments of the myostatin I gene into pGL3-basic vector and constructed plasmids pMSTN5.9K, pMSTN5.18K, pMSTN3.4K, pMSTN2.85K, pMSTN2.55K, pMSTN1.69K and pMSTN0.6K. As transfection to C2C12, ZFL (zebrafish liver cell) and NIH3T3 cells, the expression of luciferase activity was analysed. The results indicate that there were 12 E-boxes present in the 3.4Kb fragment of the zebrafish myostatin promoter, E11 and E12 E-box plays an important role in the regulation of the promoter activity in C2C12 and ZFL cells.
We utilized microinjection of an antisense RNA-expressing vector to establish a myostatin gene knockdown hereditarily stable zebrafish strain with a double-muscle phenotype. Through real time-PCR and immunostaining analysis, the myostatin messenger (m)RNA and protein levels of homozygous transgenic zebrafish were only 33% and 26% those of the non-transgenic control, respectively. And the mRNA level of myogenic regulatory factor markers, such as MyoD, myogenin, Mrf4, and Myf5 were dramatically increased in myostatin-suppressed transgenic fish compared to the non-transgenic controls. Although there was no significant difference in body length, homozygous transgenic zebrafish showed 45% heavier body weights than those of non-transgenic controls. Histochemical analysis of 1-month-old fry showed that the area of the cross-section of homozygous transgenic fish muscle fiber was twice as large as that of non-transgenic controls. This is the first model zebrafish with a hereditarily stable myostatin-suppressed genotype and a double-muscle phenotype. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T19:59:53Z (GMT). No. of bitstreams: 1 ntu-99-D89623504-1.pdf: 2142957 bytes, checksum: 019bf63f9b4fb03fe0f2a26573c4ad1e (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 論文口試委員審定書
謝辭 Ⅰ 中文摘要 Ⅱ 英文摘要 Ⅳ 目錄 Ⅴ 圖目錄 Ⅸ 表目錄 Ⅹ 第一章 文獻整理 1 1.1、肌肉倍增基因 1 1.2、Myostatin 的功能 4 1.3、肌肉生成 6 1.4、肌肉生成之調控機制 8 1.5、Myostatin 的調控 9 1.6、魚類的肌肉倍增基因 11 1.7、研究動機與目的 14 第二章 myostatin 啟動子的篩選與分析 15 2.1、摘要 15 2.2、前言 15 2.3、材料 16 2.3.1、實驗物種 16 2.3.2、實驗細胞 16 2.3.3、菌種 16 2.3.4、質體 DNA 17 2.3.5、 TA cloning 載體 17 2.3.6、 酵素 17 2.3.7、 商品化反應試劑套組 17 2.3.8、 藥品 18 2.4、方法 18 2.4.1、 斑馬魚 myostatin啟動子之篩選 18 2.4.1.1、 噬菌體宿主細胞之培養 18 2.4.1.2、 斑馬魚基因體基因庫轉印 18 2.4.1.3、 核酸探針之製備 19 2.4.1.4、 噬菌體溶菌斑雜合反應 20 2.4.2、 Lambda 噬菌體重組基因之純化 20 2.4.3、 myostatin基因體之分離與鑑定 21 2.4.4、 pMSTN-1 EGFP載體之構築 22 2.4.5、 勝任細胞製備 22 2.4.6、 質體轉型 22 2.4.7、 菌落雜合反應與顯影 23 2.4.8、 小量質體製備 23 2.4.9、 質體DNA的鑑定 24 2.4.10、 大量質體製備 24 2.4.11、 DNA序列定序與分析 25 2.4.12、 myostatin 啟動子選殖株之轉殖 25 2.4.13、 pGL3-Basic 報導載體之構築 25 2.4.14、 起動子活性分析 26 2.5、 結果 26 2.5.1、 斑馬魚 myostatin I啟動子選殖與分析 26 2.5.2、 Myostatin I 啟動子活性分析 28 2.6、討論 35 第三章 Antisense myostatin 轉殖基因魚之功能分析 38 3.1、摘要 38 3.2、前言 38 3.3、材料 39 3.3.1、實驗物種 39 3.3.2、菌種 39 3.3.3、質體 DNA 39 3.3.4、TA cloning 載體 40 3.3.5、酵素 40 3.3.6、商品化反應試劑套組 40 3.3.7、藥品 40 3.4、方法 41 3.4.1、斑馬魚 myostatin cDNA 之篩選 41 3.4.1.1、噬菌體宿主細胞之培養 41 3.4.1.2、斑馬魚 cDNA 基因庫轉印 41 3.4.1.3、核酸探針之製備 42 3.4.1.4、噬菌體溶菌斑雜合反應 42 3.4.2、Lambda 噬菌體重組基因之純化 43 3.4.3、myostatin cDNA之分離與鑑定 44 3.4.4、pEGFP-C1載體之構築 44 3.4.5、勝任細胞製備 45 3.4.6、質體轉型 45 3.4.7、菌落雜合反應與顯影 46 3.4.8、小量質體製備 46 3.4.9、質體 DNA 的鑑定 47 3.4.10、大量質體製備 47 3.4.11、cDNA序列定序與分析 48 3.4.12、pMSTN- EGFP斑馬魚胚胎顯微注射 48 3.4.13、斑馬魚魚卵之RNA抽取 48 3.4.14、1 step RT-PCR偵測myostatin基因及肌肉相關基因 49 3.4.15、組織切片及染色 49 3.4.16、細胞大小統計分析 50 3.4.17、電顯組織塊包埋 50 3.5、結果 51 3.5.1、Antisense myostatin 0.9K DNA 轉殖螢光魚表現 51 3.5.2、Antisense myostatin 0.9K DNA 轉殖螢光魚基因型及形態 55 3.5.3、Myostatin 定量 PCR 和 Western 分析 55 3.5.4、Myostatin 基因靜默 (silence ) 斑馬魚 60 hpf 的表現型 56 3.5.5、Myostatin 基因靜默斑馬魚肌肉生長相關基因之分析 56 3.5.6、生長速率和 myostatin 基因靜默斑馬魚成魚表現型 56 3.5.7、Myostatin 基因靜默斑馬魚肌肉細胞之影響 62 3.5.8、Myostatin 基因靜默斑馬魚成魚各組織之表現情形 66 3.5.9、Myostatin 基因靜默斑馬魚成魚背部肌肉之切片 66 3.6、討論 71 3.6.1、斑馬魚 myostatin cDNA基因篩選及 SiRNA 序列反意 DNA 構築 71 3.6.2、Antisense myostatin 0.9K DNA轉殖螢光魚肌肉相關基因分析 73 第四章 總結與討論 76 4.1、總結論 76 4.2、未來展望 77 參考文獻 79 附錄 90 圖 目 錄 Fig. 2-1. Genomic structure and organization of zebrafish myostatin genes. 27 Fig. 2-2. Zebrafish myostatin gene genomic Zg15-1 and 17-3 clones. 29 Fig. 2-3. Sequencing analysis of the myostatin I promoter. 30 Fig. 2-4. The effect of 5’-deletions on the transcriptional activity of the myostatin I promoter in three difference cell lines. 34 Fig. 3-1. The construction of vector-based RNA interference. The pEGFP C1 was fused with reverse partial zebrafish myostatin II cDNA. 52 Fig 3-2. The wild type and transgene silence myostatin line zebrafish in EGFP expression. 57 Fig. 3-3. EGFP expression of transgenic zebrafish in F2 generation at different stages. 58 Fig. 3-4. The gene expression of vector-based RNAi transgenic zebrafish in different muscle marker at 1.5 month. 59 Fig 3-5. Quantitative RT-PCR analysis of zebrafish Myostatin I and II mRNA levels in 60 hpf stage. 60 Fig. 3-6. SDS-PAGE of zebrafish myostatin and immunoblotting. 61 Fig. 3-7. Myogenesis-related gene expression analysis at 60 h post fertilization (hpf). 64 Fig. 3-8. Comparison of body weight and length of wild type and transgenic zebrafish. 65 Fig. 3-9. Photomicrography of 1 month stage zebrafish in dorsal muscle. 67 Fig. 3-10. Muscle fiber size analysis of transgenic zebrafish. Section were stained with hematoxylin and eosin. 68 Fig. 3-11. Quantitative analysis of adult distribution of zebrafish myostatin in wild type heterozygous and homozygous. 69 Fig.3-12. photomicrography of adult zebrafish in longitudinal section. 70 表 目 錄 Table. 2-1. Distribution of selected transcription response element on the zebrafish Myostatin I promoter. 33 Table 3-1. The hatch rate and EGFP expression rate of transgenic Vector-based RNAi zebrafish by microinjection. 53 Table.3-2. The hatch rate and EGFP expression rate of transgenic Vector-based RNAi zebrafish in F1 generation. 54 Table 3-3. Survival rate and deformity of myostatin suppressed transgenic zebrafish at the hatching-out stage. 63 | |
| dc.language.iso | zh-TW | |
| dc.subject | 肌肉倍增基因 | zh_TW |
| dc.subject | 斑馬魚 | zh_TW |
| dc.subject | 基因靜默 | zh_TW |
| dc.subject | 基因轉殖魚 | zh_TW |
| dc.subject | 啟動子 | zh_TW |
| dc.title | 斑馬魚 Myostatin 基因之分子調控與功能分析 | zh_TW |
| dc.title | Myostatin gene regulation and functional assay in zebrafish | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 黃健雄,潘子明,陳宗嶽,陸振岡,陳志毅,龔紘毅 | |
| dc.subject.keyword | 斑馬魚,肌肉倍增基因,啟動子,基因靜默,基因轉殖魚, | zh_TW |
| dc.subject.keyword | zebrafish,myostatin,gene silence,promoter,transgene fish, | en |
| dc.relation.page | 97 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2010-02-11 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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