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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.author | Yih Sheau-Wen | en |
dc.contributor.author | 易曉雯 | zh_TW |
dc.date.accessioned | 2021-07-01T08:14:13Z | - |
dc.date.available | 2021-07-01T08:14:13Z | - |
dc.date.issued | 1987 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75618 | - |
dc.description.abstract | 近年來,對於Rat貯精囊分泌蛋白的研究,已知其含有5個主要蛋白,有關其生物功能,目前尚不清楚。本實驗綜合硫銨?析、離子交換層析及再層析,來純化Mouse SVS,經由SDS-PAGE電泳分析其純度,發現貯精囊的分泌蛋白以SVS-Ⅳ含量最多。由胺基酸分析,其多勝鏈的分子量為10 Kd,含有多量的絲胺酸,缺少色胺酸及半胱胺酸,與Rat SVS-Ⅳ有許多性質相似,比較二者胺基酸次序,發現有60%以上的同源關係。經CD spectropolarimeter分析,2個蛋白均以不規則的二級結構存在。 過去的結果已知鑭系元素可當成Ca+2的替代物,藉此來研究蛋白分子是否具有Ca+2-binding sites。利用螢光分析,發現SVS-Ⅳ307 nm螢光減少及Tb+3 486nm螢光增強的現象,來研究Tb+3與SVS-Ⅳ的結合。在中性溶液中,Tb+3和mSVS-Ⅳ, rSVS-Ⅳ的結合常數分別是(2.45士0.19)x103M-1, (9.4±0.93)x102M-1。改變溶液pH值,Tb+3與SVS-Ⅳ的結合常數並未改變,但SVS-Ⅳ的立體結構可能已發生變化。 由CD光譜的研究可知,Ca+2對SVS-Ⅳ的二級結構沒有影響,且Ca+2也無法取代Tb+3-SVS-Ⅳ之位置,由此推斷SVS-Ⅳ,可能沒有Ca+2鍵結的位置。另外,SVS-Ⅳ經證明為非磷化蛋白,且mSVS-Ⅳ不具有醣分子。 台灣雨傘節(Bungarus multicintus)毒液中之β-Bungaro-toxin是由兩條多勝鏈(A, Bchains)以雙硫鍵連接。抗血清經親和層析純化得到的抗體,可同時與分開的A, B鏈反應,對其他鍵前神經毒素如:Caudoxin、Crotoxin、Mojave toxin、及Taipoxin則無反應,利用此專一性的抗體,綜合Streptavidin-biotin system,發展酵素免疫分析,提高測定的靈敏度。 | zh_TW |
dc.description.abstract | The secretory protein IV (SVS-IV) was purified from the seminal vesicle of mouse by a combination of ammonium sulfate fractionation, ion exchange chromatography to yield a product that appeared homogeneous in SDS polyacrylamide gel electrophoresis. SVS-IV is the most abundant protein in the seminal vesicle secretions. From the amino acid sequence, the molecular weight is about l0Kd; it lacks tryptophan and cysteine; serine comprises a relatively large proportion of this protein. There are similar properties in rat SVS-IV, too. Furthermore, mouse SVS-IV amino acid sequence has 60% homology with rat SVS-IV. The circular dichroic spectra indicated the absence of ordered secondary structure in the SVS-IV molecule. The interactions of terbium and SVS-IV were investigated by enhancement of Tb(III) fluorescence (λex280nm, λem486nm) and decrease in the tyrosine fluorescence(λex280nm,λem307nm). The association constants of Tb(III) to mouse SVS-IV and to rat SVS-IV were determined to be about (2.45±0.19)x103 M-1 and (9.4±0.93)x102 M-1, respectively. In line with the pH dependence of Tb(III)-SVS-IV complex formation and with the constant of binding affinity as Tb(III) bound to SVS-IV at pH 6.5, 7.0 and 7.5, we suspected that pH-induced conformational change in the protein molecule was accompanied by an increase of the energy transfer efficiency. The stepwise addition of Ca(II) to Tb(III)-SVZ-IV solution didn’t yield a large replacement of tyrosine- sensitized Tb(III) fluorescence enhancement. From the results of terbium emission, we believed that rSVS-IV and mSJS-IV had no Ca++-binding site. Quantitation of phosphate suggested that SVS-IV existed in the non-phosphorylated form before its release from the gland lumen. The mouse SVS-IV antiserum was highly specific, which didn’t show cross-reaction with rat SVS-IV. β-Bungarotoxin, a presynaptically active polypeptide, consists of two subunits which are linked by S-S bridges. We had prepared β-BuTx antibody which bound specifically to either subunit. The antibody gave very low binding to both non-neurotoxin phospholipase A2 and other presynaptic neurotoxins such as Caudoxin, Crotoxin, Mojave toxin and Taipoxin. Using the antibody, we had developed an enzyme immunoassay which could detect β-BuTx to an amount as low as 15 pg. | en |
dc.description.provenance | Made available in DSpace on 2021-07-01T08:14:13Z (GMT). No. of bitstreams: 0 Previous issue date: 1987 | en |
dc.description.tableofcontents | 1 縮寫表----------------------------3 2 中文摘要(一)----------------------5 中文摘要(二)----------------------7 第一部分: 1. 緒言------------------------------8 2. 材料、藥品、儀器------------------12 3. 實驗方法--------------------------13 4. 結果------------------------------17 5. 討論------------------------------39 第二部分: 1. 緒言------------------------------42 2. 材料、藥品------------------------44 3. 實驗方法 -------------------------45 4. 結果 -----------------------------48 5. 討論------------------------------55 附錄 英文摘要 ---------------------------58 誌謝 -------------------------------61 參考文獻----------------------------62 | |
dc.language.iso | zh-TW | |
dc.title | (一)囓齒類貯精囊分泌蛋白( SVS-Ⅳ)的純化及研究 (二)β-雨傘節毒素的酵素免疫分折 | zh_TW |
dc.title | Purification and Characterization of an Androgen-dependent Seminal Vesicle Secetory Protein Ⅳ from Rodents Enzyme Immunoassay of β-Bungarotoxin | en |
dc.date.schoolyear | 75-2 | |
dc.description.degree | 碩士 | |
dc.relation.page | 68 | |
dc.rights.note | 未授權 | |
dc.contributor.author-dept | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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