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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Jo-Wen Liu | en |
| dc.contributor.author | 劉若文 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:13:39Z | - |
| dc.date.available | 2021-07-01T08:13:39Z | - |
| dc.date.issued | 1985 | |
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W. Tsui and D. Crews. (1977). Evolution of gonadotropin structure and function. Recent Progr. Hormone Res. 33,169-248. Lowery, O. H., N. T. Rosebroug., A. L. Farrand and R. J. Randall. (1951).Protein measurement with phenol reagent. J. Biol. Chem. 193,265-275. Morgan. F. J., R. E. Canfield., J. L. Vaitukaitis and G. T. Ross. (1974). Properties of the subunits of human chorionic gonadotropin. Endocrinology 94,1601-1606. Moyle, W. R. and J. Ramachandran. (1973). Effect of LH on steroidgenesis and cyclic AMP accumulation in rat Leydig cell preparation and mouse Leydig cells. Endocrinology 93,127-134. Papkoff, H. (1974).Chemical and biological properties of the subunits of pregnant mare serum gonadotropin. Biochem. Biophys. Res. Commun. 58,397-405. Papkoff, H. and M. Ekblad. (1970).Ovine follicle stimulating hormone: Preparation and characterization of its subunits. Biochem. Biophys. Res. Commun. 40,414-621. Papkoff, H., S. W. Farmer and P. Licht. (1976).Isolation and characterization of follicle stimulating hormone and luteinizing hormone and its subunis from snapping turtle (Chelydra serpentina) pituitaries. Endocrinology 98, 767-777. Papkoff, H., M. R. Sairam and C. H. Li. (1971). The amino acid sequence of the subunits of ovine interstitial cell-stimulating hormone. J. Amer. Chem. Soc. 93, 1531. Pierce, J. G., M. R. Faith and E. M. Donaldson. (1976). Antibodies to reduced S-carboxymethylated alpha subunit of bovine luteinizing hormone and their application to study of the purification of gonadotropin from salmon (Oncorhynchus tshawtscha) pituitary. Gen. Comp. Endocrinolo. 30, 47-60. Pierce, J. G. and T. F. Parsons. (1981).Glycoprotein hormones: structure and function. Ann. Rev. Biochem. 50,465-495. Ranta, T., H. C. Chen., Y. Shimohigashi., A. J. Baukal., N. Knecht and K. J. Catt. (1985). Enhanced follicle-stimulating hormone activity of deglycosylated human chorionic gonadotropin in ovarian granulosa cells. Endocrinology 116, 59-64. Rathnam, P. and B. B. Saxena. (1971).Subunits of luteinizing hormone from human pituitary glands. J. Biol. Chem. 246, 7087-7094. Reichert, Jr. L. E. and D. N. Ward. (1974). On the isolation and characterization of the alpha and beta subunits of human pituitary follicle-stimulating hormone. Endocrinology 94, 655-664. Swaminathan. N. and O. P. Bahl. (1970). Dissociation and recombination of the subunits of human chorionic gonadotropin. Biochem. Biophys. Res. Commun. 40,422-427. Swann, R. W. and C. H. Li. (1980).Isolation and characterization of –endophin-like peptides from bovine brains. Proc. Natl. Acad. Sci. USA.77, 230-233. Tsuruhar, T., M. L. Dufan., J. Hickman and K. J. Catt. (1972). Biological properties of hCG after removal of terminal sialic acid and galactose residues. Endocrinology 91, 296-301. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75520 | - |
| dc.description.abstract | 本實驗的目的是分離鯉魚腦下腺性腺刺激素的次單元並對各次單元進行化學特性及生物活性的探討。 鯉魚腦下腺以40％乙醇，6％醋酸銨PH5.1的溶液於4℃下萃取，再以乙醇（最後濃度80%）沉澱醣蛋白。所得之醣蛋白以Sephadex G-25膠質管柱層析去除?類，之後進行DEAE-Cellulose離子交換管柱層析，收集120?300mM碳酸氫銨所條洗的部分，稱為DE300。DE300再以Sephadex G-100膠質管柱層析，得到五個部分，其中第二部分(FII)具有最強的性腺刺激素活性，此部分即本實驗所稱的鯉魚性腺刺激素。 本實驗以高壓液相層析分離鯉魚性腺刺激素各次單元，操作條件如下：分離管柱為Chemcopak7 ODS-H (4.6×250mm)之逆相管，溶劑系統包括溶劑A(水：氰甲烷?三氟醋酸＝80:20:0.04體積比)及溶劑B(水：甲醇?氰甲烷?三氟醋酸＝62.5:12.5:25:0.04體積比)；滌洗條件為?溶劑B對溶劑A作線性梯度的濃度增加，於28分鐘時，溶劑B達50％。 cGTH(FII)在此系統中展開為9個部分，依其滯留時間的短長，分稱F1?F9。經過氨基酸組成分析，部分N-末端氨基酸順序解析，以及各次單元重組結合生物活性測定，我們認為F1?F5屬於性腺刺激素一個次單元而F6?F9屬於另一次單元。F1?F9的醣類組成也被測定。 鯉魚性腺刺激素與羊黃體成長激素的次單元重組的實驗發現，在四種可能的組合中，只有羊LHα與鯉F8的重組體對促進鯉魚睪丸組織雄性素的產生具有協同作用，而其他三種組合則無，故我們認為F1?F5相當於哺乳類的α次單元而F6?F9則對應於β次單元。 | zh_TW |
| dc.description.abstract | The present study was aimed to isolate the subunits of carp gonadotropin (cGTH) and then to characterize their. chemical and biological properties. Carp pituitary glands were extracted with 40% ethanol-6% ammonium acetate, PH 5.1 at 4℃. The crude glycoprotein was desalted by Sephadex G-25 gel filtration and then chromatographed by DEAE-cellulose. Carp GTH was eluted by 120-300 mM NH4HCO3. This fraction was then chromato-graphed by Sephadex G-100. Five fractions were obtained, in which fraction II (FII) possessed the highest GTH activity. FII was used as intact cGTH throughout this study. In this study, isolation of subunit of cGTH was performed by high pressure liquid chromatography (HPLC). Operation was performed as follows: column, Chemcopak 7 ODS-H (4.6×250 mm); solvent system, solvent A (water: acetonitrile: trifluroacetic acid=80:20:0.04, v/v) and solvent B (water: methanol: acetonitrile: trifluroacetic acid 62.5:12.5:25:0.04, v/v); elution, concentration of solvent B increased to 50% linearly within 28 min. There were 9 fractions obtained from HPLC. They were designated as F1 to F9 according to their retention time (from short to long duration). From the data of amino acid composition, partial N-terminal sequence and the GTH activity of recombinants, we inferred that F1-F5 belonged to one subunit whereas F6-F9 belonged to another subunit. In this study, carbohydrate composition was also measured. In this study, recombination of the subunits of ovine LH and cGTH was also performed. Among the 4 possible combinations, only the recombinant of ovine LH -carp F8 possessed the synergistic effect on stimulation of carp testis to produce androgens whereas others did not. Therefore we assumed that F1-F5 and F6-F9 should correspond to mammalian α and β subunits, respectively. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:13:39Z (GMT). No. of bitstreams: 0 Previous issue date: 1985 | en |
| dc.description.tableofcontents | 摘要……………1 緒言…………3 材料與方法 一、化學藥品………7 二、重要儀器……………7 三、鯉魚腦下腺之來源………………8 四、性腺刺激素之純化 1．萃取粗醣蛋白……………………9 2．離子交換色層分析………………9 3．膠質滲透色層分析………………10 五、性腺刺激素生物活性測定 1．組織之製備及培育…………………10 2．固酮類激素的萃取…………………11 3．利用HPLC解析固酮激素…………………12 六、利用HPLC分離性腺刺激素之次單元………16 七、化學特性之分析 1．氨基酸組成分析…………16 2. N-末端氨基酸分析…………17 3．醣類成份分析……………18 八、性腺刺激素次單元的互相結合 1．鯉魚性腺刺激素次單元的結合………22 2．羊的黃體成長激素次單元與鯉魚性腺刺激素次單元的結合………22 九、性腺刺激素各次單元及次單元結合體之生物活性測定 1．鯉魚睪丸組織離體生成雄性素…………22 2．大白鼠萊氏細胞離體生成睪固酮……23 結果 一、鯉魚性腺刺激素之分離…………………26 二、HPLC 分離性腺刺激素次單元…………26 三、各次單元的化學特性……………………30 四、鯉魚腦下腺性腺刺激素各次單元及次單元結合體的生物活性………36 五、羊的黃體成長激素與鯉魚性腺刺激素次單元結合體的生物活性……44 討論…………………49 英文摘要………………55 謝辭…………………57 參考文獻……………58 | |
| dc.language.iso | zh-TW | |
| dc.title | 鯉魚腦下腺性腺刺激素次單元之分離及特性研究 | zh_TW |
| dc.title | Isolation and Characterization of Subunits of Carp Pituitary Gonadotropin | en |
| dc.date.schoolyear | 73-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 66 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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