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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 蔡懷楨(Huai-Jen Tsai) | |
| dc.contributor.author | Yen-Ting Chen | en |
| dc.contributor.author | 陳彥廷 | zh_TW |
| dc.date.accessioned | 2021-06-08T00:08:21Z | - |
| dc.date.copyright | 2013-08-28 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-12 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17353 | - |
| dc.description.abstract | 與綠色螢光蛋白質(green fluorescent protein, GFP)相似之GFP-like protein fam-ily的成員都具有類似的蛋白質結構，其中三個胺基酸X-Tyr-Gly會形成特殊的chromophore結構，這chromophore使GFP-like proteins具有螢光或是色澤的特性。而不同的光譜特性包括最大吸收波、最大激發波、最大發散波、量子產率(quantum yield, φ)及吸收係數(extinction coefficient, ε)，是各種GFP-like proteins的辨識方法。GFP-like proteins中最普遍的為綠色螢光蛋白質、紅色螢光蛋白質(red fluorescent protein, RFP)和不具螢光特性之色澤蛋白質(chromoprotein)，這些蛋白質的chromophore構造及族譜(protein phylogeny)已大致被分析，但種類仍然有限。目前所知的發散波在460 nm附近之螢光蛋白質皆是由綠色螢光蛋白質或紅色螢光蛋白質突變而來，且有亮度較低及穩定性不佳等缺點。本實驗室先前由海洋生物分離出紫色色澤蛋白質，最大吸收波為578 nm，經過點突變法(point mutation)後將Glu-63突變成Leu，所得之突變蛋白質之最大吸收波會偏移至516 nm，而具有最大發散波在461 nm的螢光，但螢光強度仍然偏低。
本實驗利用UV光刺激該突變蛋白質，可觀察到其最大吸收波紅移至573 nm且螢光性質消失。為了穩定及加強其螢光性質，進一步將Tyr-64突變成Leu，則形成相對較穩定的螢光蛋白質，其最大吸收波偏移至412 nm，最大發散波長為458 nm， quantum yield為0.79。若將Tyr-64突變成His和Phe，則最大放射波長分別偏移至451 nm和460 nm，quantum yield分別為0.17和0.35。軟體模擬計算出距離chromophore結構5 A內之胺基酸，將其突變成利於chromophore結構之穩定的突變蛋白質。這些蛋白質之最大放射波長皆維持在460 nm附近，其中63L/64L/90F、63L/64L/145D和63L/64L/158Q的螢光有稍微減弱之趨勢，quantum yield分別為0.60、0.66和0.73；但另外兩突變型蛋白質的螢光強度提升，quantum yield分別為0.91和0.95。這些新型的螢光蛋白質，無論在大腸桿菌(in vitro)或斑馬魚(in vivo)體內都會表現螢光。藉由改變蛋白質上的胺基酸，直接地或間接地改變chromophore的構形和鍵結，進而改變GFP-like proteins的光譜特性，而找到由shCP突變出具有高quantum yield且穩定的螢光蛋白質。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-08T00:08:21Z (GMT). No. of bitstreams: 1 ntu-102-R00b43010-1.pdf: 1852058 bytes, checksum: 15d56bf783f698d7ec1790c1f570f35b (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 中文摘要 1
英文摘要 3 文獻回顧 5 前言 16 實驗材料與方法 19 結果 27 討論 35 參考文獻 45 圖表 56 | |
| dc.language.iso | zh-TW | |
| dc.title | 具高量子產率之新型突變蛋白質 | zh_TW |
| dc.title | Novel Mutated Proteins with High Quantum Yield | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 宣大衛(David Shiuan),嚴宏洋(Hong-Young Yan),楊啟伸(Chii-Shen Yang) | |
| dc.subject.keyword | 螢光蛋白質,色澤蛋白質,基因轉殖,蛋白質突變, | zh_TW |
| dc.subject.keyword | fluorescent protein,chromoprotein,transgenic,protein mutagenesis, | en |
| dc.relation.page | 76 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2013-08-12 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
| Appears in Collections: | 分子與細胞生物學研究所 | |
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