Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/89171
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dc.contributor曾夢蛟zh_TW
dc.contributor.authorJia-Yu Leeen_US
dc.contributor.author李家宇zh_TW
dc.contributor.other園藝學系所zh_TW
dc.date2015zh_TW
dc.date.accessioned2015-12-07T06:29:14Z-
dc.identifierU0005-2008201516442000zh_TW
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Hartmann, andA. Liu. 2014. Production of functional native human interleukin-2 in tobacco chloroplasts.Mol. Biotechnol. 56(4): 369-376. Zou, Z., C. Eibl, and K. HU. 2003 The stem-loop region of the tobacco psbA 5'UTR is an important determinant of mRNA stability and translation efficiency. Mol. Genet. Genomics 269: 340-349.zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/89171-
dc.description.abstractRice(Oryza sativa L.) is one of the most important crops grown worldwide, and also is the staple food and vital crop for the inhabitants in Taiwan. Currently, gene transformation offers new possibilities for obtaining high yielding, better quality rice cultivars, and also those resistant to major diseases. Using transgenic crops as bioreactors to produce industrial, feed and fodder additives, and pharmaceutical proteins become a new way for increasing economic values of crops and may solve the problems we are facing. Tetra-peptide, VVYP, is known to inhibit fat absorption in the digestive tract and to enhance the activity of hepatic triglyceride lipase to increase the clearance of body fat. Expression of foreign genes via chloroplast genomes offers several unique advantages, including high protein levels, expressing multiple genes in operons, transgene containment by maternal inheritance, as well as lack of gene silencing, position and pleiotropic effects. In this study, Gy5-9VVYP, βC-7VVYP and15VVYP genes are transformed into therice chloroplast by using daao/aadA gene as selectable marker gene. The objective of this study is to develop the rice chloroplast as a bioreactor for production of functional VVYP peptide with hypotriglyceridemic action. Chloroplast transformation vectors harboring the VVYP and daao/aadA genes had been constructed and transferred into the rice chloroplast via biolistic bombardment. The regenerated transplastomic plantlets were selected by D-alanine and spectinomycin. The results of PCR and RT-PCR analysis of T0 and T1 leaves indicated that the transformed genes are present in the chloroplast genome of transplastomic rice plants, and expressed its mRNA.en_US
dc.description.abstract水稻是世界重要經濟農作物,也是台灣最重要的糧食作物,有其重要的民生及經濟地位。過去五十年來,傳統育種工作對改良水稻品質與產量已有許多成效。應用生物科技轉移特定基因,可突破物種間雜交障礙,發展水稻基因轉殖技術,將可為改良水稻品系提供一嶄新新途徑。利用基因轉殖作物為生物反應器以生產工業、食品、飼料及醫藥方面的產品,在高附加價值、低成本及環保因素考量之下,是目前先進國家生物科技公司積極研究發展的重要項目。台灣土地面積及天然資源有限,因此極適合朝植物生物反應器的方向發展。因此本研究開發水稻做為生物反應器的系統,增加種植水稻的保健功效及附加經濟價值,以提昇台灣水稻產業的競爭力。 VVYP胜肽(Val-Val-Tyr-Pro) 能抑制消化道對於脂肪的吸收,以及增強肝臟中三酸甘油脂水解酵素的活性,因此能快速清理體內脂肪以達到降血脂的效果。植物葉綠體基因轉殖具有:增加轉殖基因大量表現,不會造成基因污染、基因靜寂及插入位置效應,與較細胞核基因轉移穩定等優點;因此開發葉綠體基因轉殖技術為近代生物技術的主力研發工作。 本研究計畫以daao及aadA (對照)基因作為篩選標誌基因,轉移經改造過含多套組VVYP之大豆儲藏性蛋白基因(Gy5-9VVYP、7Sb-7VVYP、15VVYP)至水稻葉綠體中。本研究之目的為探討利用水稻葉綠體為非耐抗生素篩選標誌基因的生物反應器,大量生產降血脂機能性胜肽VVYP的可行性,以提高水稻之保健功效及附加經濟價值。本研究已完成構築攜帶有多套VVYP胜肽為目標基因(Gy5-9VVYP、7Sb-7VVYP、15VVYP),daao/aadA為篩選標誌基因之水稻葉綠體轉殖載體,並以基因槍法將所構築之基因轉殖到水稻葉綠體葉片。以D-alanine或spectinomycin篩選轉殖培殖體並誘導再生成水稻植株。轉殖再生植株T0及T1葉片之PCR及RT-PCR分析之結果顯示,轉殖之改造VVYP基因已存在於轉殖及之葉綠體基因組,並表現其mRNA。zh_TW
dc.description.tableofcontents中文摘要…………………………………………………………………………………i 英文摘要………………………………………………………………………………ii 目次……………………………………………………………………………………iii 圖目次…………………………………………………………………………………vi 前言……………………………………………………………………………………1 前人研究………………………………………………………………………………3 一、活性胜肽-VVYP……………………………………………………………3 二、D型胺基酸氧化酵素……………………………………………………3 三、葉綠體基因轉殖………………………………………………………4 四、葉綠體作為生物反應器……………………………………………………6 材料與方法……………………………………………………………………………8 結果……………………………………………………………………………………15 一、 帶有改造9套VVYP-Gy5、7套VVYP-7Sb 大豆蛋白基因及15套VVYP基因為目標基因,daao或aadA為篩選標誌基因,之水稻葉綠體 轉殖載體之構築……………………………………………………………15 二、誘導水稻癒傷組織、基因槍轉殖、篩選及再生水稻植株………………20 三、轉殖再生水稻T0、T1植株之基因分析……………………………………20 討論……………………………………………………………………………………65 參考文獻………………………………………………………………………………67zh_TW
dc.language.isozh_TWzh_TW
dc.rights同意授權瀏覽/列印電子全文服務,2018-08-25起公開。zh_TW
dc.subject水稻zh_TW
dc.subject葉綠體基因轉移zh_TW
dc.subjectVVYP胜肽zh_TW
dc.subject降血脂zh_TW
dc.subjectRiceen_US
dc.subjectChloroplast Gene Transformationen_US
dc.subjectVVYP Peptideen_US
dc.subjectHypotriglyceridemic Actionen_US
dc.title含降血脂機能性胜肽VVYP基因轉殖至水稻葉綠體之研究zh_TW
dc.titleStudies on Transformation of Functional Peptide VVYP Genes with Hypotriglyceridemic Action into Transplastomic Riceen_US
dc.typeThesis and Dissertationen_US
dc.date.paperformatopenaccess2018-08-25zh_TW
dc.date.openaccess2018-08-25-
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