跳到主要內容

臺灣博碩士論文加值系統

(18.208.126.232) 您好!臺灣時間:2022/08/12 01:54
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:王怡婷
研究生(外文):Yi-Teen Wang
論文名稱:番鴨性別決定候選基因DMRT1之選殖
論文名稱(外文):In Silico and Molecular Cloning ofMuscovy Sex-determining Candidate Gene DMRT1
指導教授:薛佑玲
指導教授(外文):ylshiue
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物醫學科學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:89
中文關鍵詞:矽晶選殖分子選殖番鴨性別決定
外文關鍵詞:molecluar cloningIn silico cloningsex-determinationMuscovyDMRT1
相關次數:
  • 被引用被引用:0
  • 點閱點閱:221
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0

生產鴨用或肉用雄性番鴨,係為農牧業的重要經濟目標之一。儘管目前對性別決定機制迄今尚未闡明,操縱家禽性別決定基因,將可供發展選性繁殖系統之用。果蠅DSX、線蟲MAB-3基因與人類、鼠科動物、雞隻、魚類、海龜與短吻鱷性別決定基因DMRT1,在演化上顯示顯示具高度保留性。因此,雞隻性染色體Z連鎖之DMRT1基因,成為家禽調控性別分化之候選調節基因。本論文目的要選殖並瞭解番鴨DMRT1基因,以進一步應用於選性繁殖上。目前以定序出番鴨Muscovy DMRT1 cDNA部分序列,並顯示番鴨DMRT1 cDNA序列與雞隻及海龜分別有95%及83%相同度,而番鴨DMRT1胺基酸序列則與雞隻及海龜分別有61%及54%相同度。以Phylip程式分析各物種間之DMRT1異物種同源基因並以MEGA 2程式繪製親緣樹。以上結果顯示家禽DMRT1基因在cDNA序列與胺基酸序列層次上之親緣關係,均與爬蟲類較相近,而與哺乳類有較遠的親緣關係。


To produce male Muscovy only for fatty liver and meat-type production is an important economic goal in animal husbandry, although the sex-determining mechanism in poultry remains to be elucidated. Manipulation of sex-determining gene(s) in poultry provides enormous opportunities on the development of sex pre-selection reproductive systems. DSX and MAB-3 genes in Drosophila and C. elegans are conserved across the human, mice, chickens, fish, turtles, and reptiles revealing an ancient sex-determining locus DMRT1. Thus the Z-linked, DMRT1 in chicken is an excellent candidate regulatory gene controlling similar aspects of sexual development in poultry. This dissertation is aimed to clone and characterize Muscovy DMRT1 gene for further application in sex pre-selection. Partial cDNA sequences of Muscovy DMRT1 was determined and revealed 95% identity and 83% with chicken and red-eared slider turtle DMRT1 cDNA sequences. DMRT1 orthologs among various species were analyzed by Phylip program and phylogenetic tree was constructed by MEGA2 programs. Results indicated that Muscovy, chicken and red-eared slider turtle DMRT1 revealing 95%, and 83% identity at cDNA and 61%, 54% identity at amino acid level.


中文摘要.....................................................................................................Ⅰ
英文摘要.....................................................................................................Ⅱ
英文縮寫表.................................................................................................Ⅲ
論文正文
第一章序言.............................................................................................1
第二章文獻檢討
2.1 家禽的選性繁殖................................................................3
2.2 鳥類之基因組....................................................................3
2.3 性染色體............................................................................4
2.3.1 哺乳類性染色體......................................................5
2.3.2 鳥類的性染色體......................................................5
2.3.3 鳥類性染色體與性別決定基因的演化..................6
2.4 性別決定的假說................................................................6
2.4.1 遺傳因子平衡模式..................................................7
2.4.2 顯性基因模式.........................................................7
2.5 家禽之早期性別鑑定.........................................................8
2.6 參與家禽性腺發育之基因.................................................9
2.7 W性染色體連鎖基因,Wpkci..........................................11
2.8 DMRT1
2.8.1 DMRT1異物種同源基因.......................................12
2.8.1.1線蟲MAB-3與果蠅DSX ....................................13
2.8.1.2 人類DMRT1.......................................................13
2.8.1.3 雞隻DMRT1.......................................................14
2.8.2 人類9p 缺失與DMRT1......................................14
2.8.3 比較基因定位.......................................................15
2.8.4 人類DMRT1 基因之基因結構............................15
2.9 溫度調控爬蟲類之性別決定..........................................16
2.10 番鴨................................................................................17
第三章材料與方法
3.1 矽晶選殖番鴨性別決定候選基因DMRT1.....................18
3.1.1 由GenBank 與表現序列標幟資料庫搜尋番鴨
DMRT1的異物種同源基因................................18
3.1.2 引子設計...............................................................18
3.2 分子選殖番鴨DMRT1 基因
3.2.1 番鴨.......................................................................19
3.2.2 萃取睪丸組織的total RNA..................................20
3.2.3 反轉錄-聚合? 鏈鎖反應......................................20
3.2.4 熱啟動聚合? 鏈鎖反應........................................21
3.2.5 TA cloning 與大腸桿菌轉殖作用.........................22
3.2.6 自瓊脂凝膠純化DNA 片段................................23
3.2.7 5' Rapid Amplification of cDNA Ends..................24
3.2.8 3’ Rapid Amplification of cDNA Ends..................25
3.2.9 定序.......................................................................26
3.2.10 序列分析與二級結構預測.................................27
第四章結果
4.1 矽晶選殖番鴨性別決定候選基因DMRT1.....................30
4.2 分子選殖DMRT1.............................................................31
4.3 DMRT1異物種同源基因之親緣樹..................................32
4.4 DMRT1 之二級結構預測.................................................33
第五章討論...........................................................................................34
第六章結論...........................................................................................37
參考文獻.....................................................................................................38

表一、不同物種性別發育過程中,基因型與外表型間的相對關係.....48
表二、番鴨的生活習性...........................................................................49
表三、PCR 反應所使用之引子對與PCR 反應之進行條件..................50
表四、雞隻DMRT1DNA 序列,經由blastn 檢索nr database,未發現
其他鳥類的DMRT1 異物種同源基因.......................................51
表五、以雞隻DMRT1 基因(Acc#AF123456)當作探針,所得異物種
同源序列之長度、一致性區域及百分比...................................52
表六、由資料庫所檢索DMRT1 異物種同源序列之長度、轉譯區,與
雞隻DMRT1 蛋白質序列間的相同度與相似性.......................53
表七、多重序列組合所分析之DMRT1 異物種同源基因列表...........54
表八、不同物種間DMRT1 胺基酸序列之相似性與相同度...............55

圖一、雞隻染色體Z、W 上的基因,與人類第五、八、九、十八體
染色體的異物種同源基因.........................................................56
圖二、參與脊椎動物生殖腺發育特異階段之基因..............................57
圖三、Wpkci 與雞隻PKCI(chPKCI)之胺基酸序列比對.....................58
圖四、DM domain 之圖示(a)平面(b)立體帶狀結構..........................59
圖五、DMRT1 之基因結構..................................................................60
圖六、DMRT1異物種同源基因的生物資訊研究................................61
圖七、選殖番鴨DMRT1 基因之實驗流程圖..................................62
圖八、人類與雞DMRT1 cDNA 序列的保留性區域.........................63
圖九、番鴨DMRT1之PCR產物與雞隻DMRT1間之相對位置.........64
圖十、PCR 產物A、D、H、I、N之電泳分析圖.............................65
圖十一、番鴨DMRT1 基因列之核? 酸及蛋白質序列.....................66
圖十二、以Phylip 程式比較不同物種之DMRT1 胺基酸序列.........67
圖十三、不同物種間DMRT1基因序列之親緣樹
(a)ME (b)MP........................................................................71
(c)NJ (d)UPGMA................................................................72
圖十四、DM domain 之演化高度保留性.............................................73
附錄.............................................................................................................74


參考文獻戴謙、李善男。1995。台灣省畜產試驗所家畜人工生殖技術研究之成果及未來展望。畜產研究, 28: 1-7。洪福隆、田正德、陳明造。1995。家飼番鴨的歷史(一)。中國畜牧會誌, 27:131-136。洪福隆、田正德、陳明造。1996a。家飼番鴨的歷史(七)。中國畜牧會誌, 28:71-73。洪福隆、田正德、陳明造。1996b。家飼番鴨的歷史(八)。中國畜牧會誌, 28:92-95。Baker B.S., Ridge K.A. 1980. Sex and the single cell: On the action of major loci affecting sex determination in Drosophila melanogaster. Genetics, 94: 383-423.Barbaux S., Niaudet P., Gubler M.C., Grunfeld J.P., Jaubert F., Kuttenn F., Fekete C.N., Souleyreau-Therville N., Thibaud E., Fellous M., McElreavey K. 1997. Donor splice-site mutations in WT1 are responsible for Frasier syndrome. Nature Genetics, 17: 467-470.Bloom S.E. 1981. Detection of normal and aberrant chromosomes in chicken embryos and in tumor cells. Poultry Science, 60: 1355-1361.Brunner B., Hornung U., Shan Z., Nanda I., Kondo M., Zend-Ajusch E., Haaf T., Ropers H.H., Shima A., Schmid M., Kalscheuer V.M., Schartl M. 2001. Genomic organization and expression of the doublesex-related gene cluster in vertebrates and detection of putative regulatory regions for DMRT1. Genomics, 77: 8-17.Burtis K.C., Baker B.S. 1989. Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides. Cell, 56: 997-1010.Calvari V., Bertini V., De Grandi A., Peverali G., Zuffardi O., Ferguson-Smith M., Knudtzon J., Camerino G., Borsani G., Guioli S. 2000. A new submicroscopic deletion that refines the 9p region for sex reversal. Genomics, 65: 203-212.Capel B. 1998. Sex in the 90s: SRY and the switch to the male pathway. Annual Review in Physiology, 60: 497-523.Capel B. 2000. The battle of the sexes. Mechanisms of Development, 92: 89-103. Christidis L. 1990. Chordata 3. B. Aves. Animal cytogenetics. Volume IV, pp. 1-108. Gebruder Borntraeger Verlagsbuchhandlung, Science Publishers, Berlin, Germany. Clinton M. 1996/1997. Sex determination in birds. Roslin Institute, Edinburgh, Annual Report, 55. Clinton M. 1998. Sex determination and gonadal development: a bird's eye view. Journal of Experimental Zoology, 281: 457-465.Clinton M., Haines L.C. 1999. An overview of factors influencing sex determination and gonadal development in birds. Cellular and Molecular Life Sciences, 55: 876-886.David H.E., George F.G., and Claire M.M., 1996. Cranes: their biology, husbandry and conservation. pp. 223-230. Department of the Interior, National Biological Service, Wisconsin. USA.Ellegren H. 2000. Evolution of the avian sex chromosomes and their role in sex determination. Trends in Ecology and Evolution, 15: 188-192.Ellegren H. 2001. Hens, cocks and avian sex determination. A quest for genes on Z or W? European Molecular Biology Organization Report, 2: 192-196.Fridolfsson A.K., Cheng H., Copeland N.G., Jenkins N.A., Liu H.C., Raudsepp T., Woodage T., Chowdhary B., Halverson J., Ellegren H. 1998. Evolution of the avian sex chromosomes from an ancestral pair of autosomes. Proceeding of the National Academy of Sciences of the United Stated of America, 95: 8147-8152.Ferguson M.W., Joanen T. 1982. Temperature of egg incubation determinessex in Alligator mississippiensis. Nature, 296: 850-853.Gregory T.R. 2001. Animal Genome Size Database. http://www.genomesize. comGriffiths R. 2000. Sex identification in birds. Seminiars in Avian and Exotic pet medicine, 9: 14-26. Guan G., Kobayashi T., Nagahama Y. 2000. Sexually dimorphic expression of two types of DM (Doublesex/Mab-3)-domain genes in a teleost fish, the Tilapia (Oreochromis niloticus). Biochemical and Biophysical Research Communications, 272: 662-666.Halverson J.L., Dvorak J. 1993. Genetic control of sex determination in birds and the potential for its manipulation. Poultry Science, 72: 890-896.Hiort O., Holterhus P.M. 2000. The molecular basis of male sexual differentiation. European Journal of Endocrinology, 142: 101-110. Hori T., Asakawa S., Itoh Y., Shimizu N., Mizuno S. 2000. Wpkci, encoding an altered form of PKCI, is conserved widely on the avian W chromosome and expressed in early female embryos: implication of its role in female sex determination. Molecular Biology of the Cell, 11: 3645-3660.Huang X., Cheng H., Guo Y., Liu L., Gui J., Zhou R. 2002. A conserved family of doublesex-related genes from fishes. Journal of Experimental Zoology, 294: 63-67.Kettlewell J.R., Raymond C.S., Zarkower D. 2000. Temperature-dependent expression of turtle Dmrt1 prior to sexual differentiation. Genesis, 26: 174-178.Kumar S., Tamura K., Jakobsen I.B., Nei M. 2001. MEGA2: molecular evolutionary genetics analysis software. Bioinformatics, 17: 1244-1245.Lim H.N., Hawkins J.R. 1998. Genetic control of gonadal differentiation. Bailliere's Clinical Endocrinology and Metabolism, 12: 1-16.Lima C.D., Klein M.G., Weinstein I.B., Hendrickson W.A. 1996. Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins. Proceeding of the National Academy of Sciences of the United Stated of America, 93: 5357-5362.Marchand O., Govoroun M., D'Cotta H., McMeel O., Lareyre J., Bernot A., Laudet V., Guiguen Y. 2000. DMRT1 expression during gonadal differentiation and spermatogenesis in the rainbow trout, Oncorhynchus mykiss. Biochemical and Biophysical Research Communications, 1493: 180-187.Marshall Graves J.A., Shetty S. 2001. Sex from W to Z: evolution of vertebrate sex chromosomes and sex determining genes. Journal of Experimental Zoology, 290: 449-462.McCarrey J. R. and Abbott U. K. 1979. Mechanisms of genetic sex determination, gonadal sex differentiation, and germ-cell development in animals. Advances in Genetics, 20: 217-290. Moniot B., Berta P., Scherer G., Sudbeck P., Poulat F. 2000. Male specific expression suggests role of DMRT1 in human sex determination. Mechanisms of Development, 91: 323-325.Nachtigal M.W., Hirokawa Y., Enyeart-VanHouten D.L., Flanagan J.N., Hammer G.D., Ingraham H.A. 1998. Wilms' tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex-specific gene expression. Cell, 93: 445-454.Nanda I., Zend-Ajusch E., Shan Z., Grutzner F., Schartl M., Burt D.W., Koehler M., Fowler V.M., Goodwin G., Schneider W.J., Mizuno S., Dechant G., Haaf T., Schmid M. 2000. Conserved synteny between the chicken Z sex chromosome and human chromosome 9 includes the male regulatory gene DMRT1: a comparative (re)view on avian sex determination. Cytogenetics and Cell Genetics, 89: 67-78.Nanda I., Shan Z., Schartl M., Burt D.W., Koehler M., Nothwang H., Grutzner F., Paton I.R., Windsor D., Dunn I., Engel W., Staeheli P., Mizuno S., Haaf T., Schmid M. 1999. 300 million years of conserved synteny between chicken Z and human chromosome 9. Nature Genetics, 21: 258-259.Nakamura D., Tiersch T.R., Douglass M., Chandler R.W. 1990. Rapid identification of sex in birds by flow cytometry. Cytogenetic and Cell Genetics, 53: 201-205.O'Neill M., Binder M., Smith C., Andrews J., Reed K., Smith M., Millar C., Lambert D., Sinclair A. 2000. ASW: a gene with conserved avian W-linkage and female specific expression in chick embryonic gonad. Development, Genes and Evolution, 210: 243-249.Ottolenghi C., Fellous M., Barbieri M., McElreavey K. 2002. Novel paralogy relations among human chromosomes support a link between the phylogeny of doublesex-related genes and the evolution of sex determination. Genomics, 79: 333-343.Ottolenghi C., McElreavey K. 2000. Deletions of 9p and the quest for a conserved mechanism of sex determination. Molecular Genetics and Metabolism, 71: 397-404.Ottolenghi C., Veitia R., Quintana-Murci L., Torchard D., Scapoli L., Souleyreau- Therville N., Beckmann J., Fellous M., McElreavey K. 2000. The region on 9p associated with 46,XY sex reversal contains several transcripts expressed in the urogenital system and a novel doublesex-related domain. Genomics, 64: 170-178.Parker K.L. 1998. The roles of steroidogenic factor 1 in endocrine development and function. Molecular and Cellular Endocrinology, 145: 15-20.Parker K.L., Schedl A., Schimmer B.P. 1999. Gene interactions in gonadal development. Annual Review in Physiology, 61: 417-433.Parker K.L., Schimmer B.P., Schedl A. 1999. Genes essential for early events in gonadal development. Cellular and Molecular Life Sciences, 55: 831-838.Parker T.R., Birkhead T.R., Joshua S.K. 1992. Sex ratio in a population of guillemots Uria aalge determined by chromosome analysis. The Ibis, 133: 423-425.Pieau C., Dorizzi M., Richard-Mercier N. 1999. Temperature-dependent sex determination and gonadal differentiation in reptiles. Cellular and Molecular Life Sciences, 55: 887-900.Raymond C.S., Shamu C.E., Shen M.M., Seifert K.J., Hirsch B., Hodgkin J., Zarkower D. 1998. Evidence for evolutionary conservation of sex-determining genes. Nature, 391: 691-695.Raymond C.S., Kettlewell J.R., Hirsch B., Bardwell V.J., Zarkower D. 1999. Expression of Dmrt1 in the genital ridge of mouse and chicken embryos suggests a role in vertebrate sexual development. Developmental Biology, 215: 208-220.Raymond C.S., Parker E.D., Kettlewell J.R., Brown L.G., Page D.C., Kusz K., Jaruzelska J., Reinberg Y., Flejter W.L., Bardwell V.J., Hirsch B., Zarkower D. 1999. A region of human chromosome 9p required for testis development contains two genes related to known sexual regulators. Human Molecular Genetics, 8: 989-996.Raymond C.S., Murphy M.W., O'Sullivan M.G., Bardwell V.J., Zarkower D. 2000. Dmrt1, a gene related to worm and fly sexual regulators is required for mammalian testis differentiation. Genes and Development, 14: 2587-2595.Richner H. 1989. Avian laparoscopy as a field technique for sexing birds and an assesment of its effects on wild birds. Journal of Field Ornithology, 60: 137-142.Saitou N., Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4: 406-425.Shan Z., Nanda I., Wang Y., Schmid M., Vortkamp A., Haaf T. 2000. Sex-specific expression of an evolutionarily conserved male regulatory gene, DMRT1, in birds. Cytogenetics and Cell Genetics, 89: 252-257.Shen M.M., Hodgkin J. 1998. mab-3, a gene required for sex-specific yolk protein expression and a male-specific lineage in C. elegans. Cell, 54: 1019-1031.Shetty S., Griffin D.K., Graves J.A. 1999. Comparative painting reveals strong chromosome homology over 80 million years of bird evolution. Chromosome Research, 7: 289-295.Smith C.A., McClive P.J., Western P.S., Reed K.J., Sinclair A.H. 1999. Conservation of a sex-determining gene. Nature, 402: 601-602.Smith C.A., Smith M.J., Sinclair A.H. 1999. Gene expression during gonadogenesis in the chicken embryo. Gene, 234: 395-402.Sneath P.H.A., Sokal R.R. 1973. in Numerical Taxonomy, pp.230-234. W.H. Freeman and Company. San Francisco. California. USASpotila L.D., Spotila J.R., Hall S.E. 1998. Sequence and expression analysis of WT1 and Sox9 in the red-eared slider turtle, Trachemys scripta. Journal of Experimental Zoology, 281: 417-427.Stevens L. 1996. Avian biochemistry and molecular biology, pp.117-118. Cambridge University Press. New York. USA.Stevens L. 1997. Sex chromosomes and sex determining mechanisms in birds. Science Progress, 80: 197-216.te Winkel G.P. 1997. Biosecurity in poultry production: where are we and where do we go? Acta Veterinaria Hungarica, 45: 361-372.Veitia R.A., Salas-Cortes L., Ottolenghi C., Pailhoux E., Cotinot C., Fellous M. 2001. Testis determination in mammals: more questions than answers. Molecular and Cellular Endocrinology, 179: 3-16.Western P.S., Harry J.L., Graves J.A., Sinclair A.H. 1999. Temperature- dependent sex determination in the American alligator: AMH precedes SOX9 expression. Developmental Dynamics: an Official Publication of the American Association of Anatomists, 216: 411-419.Yi W., Zarkower D. 1999. Similarity of DNA binding and transcriptional regulation by Caenorhabditis elegans MAB-3 and Drosophila melanogaster DSX suggests conservation of sex determining mechanisms. Development, 126: 873-881.Zhu L., Wilken J., Phillips N.B., Narendra U., Chan G., Stratton S.M., Kent S.B., Weiss M.A. 2000. Sexual dimorphism in diverse metazoans is regulated by a novel class of intertwined zinc fingers. Genes and Developemnt, 14: 1750-1764.

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top