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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75221完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | 宋欣和 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:12:16Z | - |
| dc.date.available | 2021-07-01T08:12:16Z | - |
| dc.date.issued | 2001 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75221 | - |
| dc.description.abstract | 在脊椎動物的胚胎發育中，Hedgehog（Hh）分泌性蛋白質路徑屬於一個重要的訊息傳遞鏈，負責神經管，腳，肺，皮膚，頭髮，牙齒的發育；大部份Hh訊息傳遞鏈內的成員具有高度的演化保守性，而且其功能大部份都是從果蠅研究中得到。 在果蠅翅碟中，Hh後半區的Hh產生細胞中表達產生、在Golgi中進行自我裂解、結合膽固醇成為具有的功能HhN-膽固醇分子、經目前未知的蛋白質傳輸到細胞膜上，細胞膜上的HhN-膽固醇分子再藉由Dispatched(Disp)膜蛋白轉送前半區的Hh接收細胞，啟動Hh標的基因產物如Patched（Ptc，Hh的受體）和Dpp（Decapentaplegic，果蠅的TGF-β同源基因）蛋白質的表達；Dpp型態因數向外散佈形成蛋白質濃度階梯決定翅碟細胞的細胞命運。 rotini(rti)是一個新的節向基因與負責運送傳輸蛋白質，具有70％的相同性，推測Rti的功能與細胞內蛋白質的運送傳輸（protein trafficking）有關。當在翅碟前半區rti突變區域位時，位在緊鄰前後半區界線的Ptc蛋白質表現範圍從野生型的5-6個細胞寬減少到2-3個細胞寬，但位在突變區域內較前方的野生型Hh接收細胞卻沒有Ptc表現；這顯示HhN-膽固醇分子無法通過rti突變區域被運送到較前方的野生型Hh接收細胞。rti在前半區的突變性狀與tout velu（ttv）（影響proteoglycan合成的基因）位在前半區的突變行為相似，我們推測Rti與Ttv皆參與在HhN-膽固醇分子在Hh接收細胞間的傳送。 當rti突變區域位在翅碟後半區Hh產生細胞區時，Hh蛋白質表現量下降、但不影響HhLacZ的表現量，這顯示Rti參與Hh在轉錄過程以後的製造行為。基於上述的資料，推測rti的功能之一與Hedgehog運送與製造。 | zh_TW |
| dc.description.abstract | In the embryogenesis of vertebrate, Hh signaling pathway is an important signaling pathway that is responsible for the development of neural tube, limb, lung, skin, hair, teeth; most components in Hh signaling pathway are evolutionally conserved. The functions of most components in Hh signaling pathway are derived from the studies in Drosophila. In Hh production cells, after autoprocessing and cholesterol modification in Golgi, Hh processing protein (Hh-Np) is trafficking to lipid rafts in plasma membrane by unknown factor(s). Hh is displaced from the lipid raft by Dispatched (Disp) and transported to receiving cells to activate Hh target genes, patched (ptc) and dpp (decapentaplegic). Dpp forms a protein gradient to pattern the cell fates in wing discs. rotini(rti), a novel Drosophila segment polarity gene, is about 70% identity with the potein that involves in protein trafficking. Rti may involve in protein trafficking in the cells. In rti mutant clone, ptc expression is only weakly induced about 2 or 3-row cells adjacent A-P border in the anterior compartment in wing disc. But ptc expression is not appeared in the wild type cells located on anterior to rti mutant clone. It is suggested that Hh-Np cannot be transported to the wild-type cells across the rti mutant clone. This situation is like the mutant clone of tout velu (ttv) that is involved in proteoglycan synthesis in wing disc. Both rti and ttv may be required for Hh transportation in Hh receiving cells. In rti mutant clone, Hh expression is diminished in the posterior compartment in wing disc, but Hh-LacZ expression is not affected. Our finding suggested that rti might involve in the post-translation process of Hh. According to the above results, we suggest that rti may be required for Hh production and transportation. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:12:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2001 | en |
| dc.description.tableofcontents | 中文摘要:………………………………………………………………………………………………………………………1 Abstract………………………………………………………………………………………………………………………3 Table Content…………………………………………………………………………………………………………………5 List of Tables………………………………………………………………………………………………………………10 List of Figure………………………………………………………………………………………………………………11 Introduction…………………………………………………………………………………………………………………16 Anterior-posterior axis formation in Drosophila embryo…………………………………………………………16 Wingless and Hedgehog signaling pathway……………………………………………………………………………17 Maintainence of the parasegment boundary……………………………………………………………………………18 Hh signaling pathway………………………………………………………………………………………………………18 Hh transportation from posterior compartment to anterior compartment………………………………………20 The posttranslational modification of Hh……………………………………………………………………………20 Dispatched……………………………………………………………………………………………………………………22 Heparan Sulfate Proteglycan……………………………………………………………………………………………23 The receptors of Hh………………………………………………………………………………………………………25 Model of Hh transportation………………………………………………………………………………………………27 Missing link in this model………………………………………………………………………………………………27 A novel segment polarity gene rotini…………………………………………………………………………………29 Materials and Methods……………………………………………………………………………………………………30 Fly stocks and maintenance………………………………………………………………………………………………30 The autosomal FLP-DFS technique………………………………………………………………………………………30 P-transposon induced mutagenesis screen for homozygous lethal mutation on the second chromosome…31 Germ-line clone production………………………………………………………………………………………………31 Cuticular preparation……………………………………………………………………………………………………32 IPCR……………………………………………………………………………………………………………………………32 Plasmid rescue………………………………………………………………………………………………………………33 Isolation plasmid DNA from Bacteria…………………………………………………………………………………34 Imprecise Excision…………………………………………………………………………………………………………34 Generation of recombinant clones in somatic tissues……………………………………………………………34 Fluorescence imaginal discs antibody staining……………………………………………………………………35 DEPC-distilled H20…………………………………………………………………………………………………………37 X-gal staining for embryo………………………………………………………………………………………………37 Mis-expression experiment………………………………………………………………………………………………38 Single fly PCR………………………………………………………………………………………………………………38 Result:………………………………………………………………………………………………………………………40 P-induced lethal lines with specific maternal effect……………………………………………………………40 Group 1. Germ cell lethal………………………………………………………………………………………………40 Group 2. Abnormal oogenesis……………………………………………………………………………………………40 Group 3. Maternal effect phenotype……………………………………………………………………………………41 Group 4. No maternal effect……………………………………………………………………………………………41 rotini is a novel mutation exhibiting lawn-of-denticles embryonic phenotype……………………………41 The embryonic phenotype of P1164A4……………………………………………………………………………………43 rotini is a novel segment polarity gene……………………………………………………………………………44 The lethality of is caused by P element insertion………………………………………………………………45 P1164A4 element disrupts the annotated gene………………………………………………………………………45 P1164A4 element may affect two genes…………………………………………………………………………………46 The segment polarity embryonic phenotype can be phenocopied by RNA interference of the predicted gene……………………………………………………………………………………………………………………………47 LacZ expression pattern of rtiA4 heterozygous embryos…………………………………………………………48 rotini is involved in Hh transportation……………………………………………………………………………49 rotini is required for Hh production…………………………………………………………………………………50 The ptc expression is diminished in the big clone across the A-P boredr…………………………………51 rotini acts in Hh signaling pathway…………………………………………………………………………………52 rti acts in Wg signaling pathway………………………………………………………………………………………53 Somatic clones in adult…………………………………………………………………………………………………55 Discussion:…………………………………………………………………………………………………………………56 Rti is a putative coat protein…………………………………………………………………………………………56 Rotini is required for the Hh transportation………………………………………………………………………57 Rti is required for Hh production……………………………………………………………………………………57 rotini acts in both Hh and Wg pathway………………………………………………………………………………58 Is Rti protein a common factor in cells……………………………………………………………………………59 Two modes of rti function………………………………………………………………………………………………60 1. rotini may involve in ttv-dependent PG synthesis……………………………………………………………60 2. rti may affect trafficking the cargo to membrane……………………………………………………………61 Acknowledgment………………………………………………………………………………………………………………63 Reference……………………………………………………………………………………………………………………64 | |
| dc.language.iso | zh-TW | |
| dc.title | 一個新的高基氏體蛋白質，Rotini，參與果蠅Hedgehog的製造與傳遞 | zh_TW |
| dc.title | Rotini, a novel putative Golgi protein, is required for the production and transportation of Drosophila Hedgehog | en |
| dc.date.schoolyear | 89-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 76 | |
| 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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