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研究生:羅筱文
研究生(外文):Hsiao wen Lo
論文名稱:藉由秀麗隱桿線蟲可表達多基因之特性用於表現HDV小型Delta抗原
論文名稱(外文):Expression of Small Delta Antigen of Hepatitis Delta Virus in Caenorhabditis elegans by Using Polycistronic Vectors
指導教授:羅時成羅時成引用關係
指導教授(外文):S. C. Lo
學位類別:碩士
校院名稱:長庚大學
系所名稱:基礎醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
論文頁數:55
中文關鍵詞:D型肝炎病毒秀麗隱桿線蟲小型delta抗原
外文關鍵詞:HDVC. eleganspolycistronicsmall delta antigenoperons
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多基因性操縱子(polycistronic operons)調控基因群中具有相似功能的基因並且高度保留於線蟲 (Caenorhabditis elegans),利用多基因性操縱子的特性,使其相對簡單的基因體能表現複雜的生物功能,結合線蟲固定的細胞系源 (cell lineage),利用線蟲的在發育生物學上的優異性,結合發育遺傳學與分子生物的方法,使得線蟲成為系統生物學最佳的實驗模式動物。D型肝炎病毒是必須依賴B型肝炎病毒才能完成整個生命週期,經過多年的研究,在細胞層次中確切的致病機制仍未解開。本論文中,藉由具有小型 delta 抗原的譯雙基因及譯三基因質體,製造表現小型delta 抗原的轉殖線蟲。其中,在線蟲咽喉部表達紅色螢光蛋白與小型 delta 抗原融合蛋白質的轉殖線蟲株中,發現有生長遲緩的現象。然而,在相似但含有終止密碼子於紅色螢光蛋白和小型 delta 抗原之間的轉殖線蟲株中,生長遲緩現象則較不顯著。此外利用RNA干擾抑制小型delta 抗原基因在轉殖線蟲株的表現以減緩生長遲緩及促使後代繁衍。總體而言,生長遲滯可能是由小型 delta 抗原在咽喉部對轉殖線蟲所造成的影響,引起發育遲滯的機制則尚待更進一步的研究。
In Caenorhabditis elegans, polycistronic operons have been widely contained and co-regulate genes of related functions in the operons; therefore, the simple but with biological complexity genome could be conserved. Combine with the invariant and well-known cell lineage development of C. elegans makes it a perfect platform for system biology research. Though hepatitis delta virus, a defective RNA virus that its complete life cycle relies on hepatitis B virus, has been studied for years the exact pathogenic mechanism in cellular level remains unclear. In this thesis, transgenic worms expressing bicistronic or tricistronic genes which included small delta antigen were obtained. One line of transgenic animals appeared growth retardation when DsRed fused with the small delta antigen was expressed in pharynx. However, transgenic animals contain a similar construct which was created a stop codon between DsRed at small delta antigen, appeared less significant in growth retardation. Furthermore, RNAi method to knock-down delta antigen gene in transgenic animals rescues their growth arrest and brood size. Taken together, the retardation might be resulted from small delta antigen expression in pharynx. The mechanism of causing retardation remains to be investigated.
Table of Content
指導教授推薦書 -i-
口試委員會審定書 -ii-
長庚大學碩士論文著作授權書 -iii-
Acknowledgements -iv-
中文摘要 -v-
Abstract -vi-
Table of Content -vii-
Table of Figures -ix-
Introduction - 1 -
Specific Aims - 5 -
Materials and Methods - 6 -
General methods and strains - 6 -
DNA constructs - 6 -
Transgenic worms and image screening - 7 -
RNA interference - 8 -
Phenotype assays - 9 -
Western blot analysis - 9 -
Results - 11 -
Construction and characteristics of plasmids used in this study - 11 -
Expression comparison of three tricistronic genes in C. elegans - 13 -
Developmental effect of S-HDAg expression in pharynx of C. elegans - 14 -
Discussion - 16 -
References - 19 -
Figures - 22 -
Appendix - 30 -


Table of Figures
Fig. 1 Plasmid maps and restriction digestion analysis by agarose gel electrophoresis - 22 -
Fig. 2 Comparison of different translational controllers - 24 -
Fig. 3 The effect of small delta antigen on pharynx development of C. elegans - 26 -
Fig. 3 The effect of small delta antigen on pharynx development of C. elegans - 27 -
Fig. 4 The growth and reproductive effects of Pmyo-2::gfp-icr-dsred::sd on C. elegans - 28 -
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