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dc.contributorMei-Er Chenen_US
dc.contributorRui-Bin Yangen_US
dc.contributorEn-Cheng Yangen_US
dc.contributor.advisorKuang-Hui Luen_US
dc.contributor.authorChung, Po-Hsuen_US
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dc.description.abstractGuanylyl cyclases (GCs), kind of enzyme-linked receptor proteins, play important physiological functions, including metamorphosis, olfactory organ and memory, in insects. The BdmGC-1, a receptor form membrane guanylyl cyclase of Bactrocera dosalis (Hendel), has been proved its relationship with eclosion hormone. In this study, we furtherfound that the BdmGC-1 was expressed in the B. dorsalis imaginal discs of wings, legs and compound eyes. Real-time PCR analysis revealed that new (1-day-old) puparium exhibited higher expression level during metamorphosis. Moreover, immunofloruscent staining showed that the BdmGC-1 signals were located in whole imaginal discs. Furthermore, recent studies have showed that p-21 activated protein (PAK) can regulate the mammalian GC activity. Hence, we used a 280-bp BdPAK, cloned from B. dorsalis based on PAK conserved regions, to analyze its expression and location. The results of both real-time PCR analysis and immunofloruscent staining showed that BdPAK has similar expression profiles with BdmGC-1, implying that there some kind of relationships are existed between these two proteins. In summary, BdmGC-1 is indeed expressed in the B. dorsalis wing, compound eye and leg discs and the newly pupated purarium have the higher expression level of BdmGC-1. However, functional interaction of BdmGC-1 with BdPAK has not yet been understood.en_US
dc.description.abstract鳥苷酸環化酶(guanylyl cyclase, GC)為昆蟲體內一重要受器蛋白(receptor protein),許多生理功能都與其相關,例如變態、嗅覺和記憶等。東方果實蠅(Bactrocera dorsalis (Hendel))鳥苷酸環化酶,BdmGC-1,已被證實為羽化激素(eclosion hormone)之受器蛋白,並發現其可能與昆蟲蛻皮行為有關。本研究更進一步發現BdmGC-1亦存在東方果實蠅幼蟲體內的翅成蟲盤(wing disc)、眼成蟲盤(eye disc)和足成蟲盤(leg disc)上。以real-time PCR 分析發現,蛹期第一天之翅、眼和足成蟲盤均有較高的BdmGC-1 表現量;以免疫螢光染色(immunofluorescent stain)分析BdmGC-1 在翅、眼和足成蟲盤表現位置,結果顯示其分布位於整個成蟲盤上。另外,從前人研究已知p-21 activated protein(PAK)在哺乳動物上可調節GC 活性。因此,以PAK 保守區域選殖得到一長度280 bp 的東方果實蠅部分PAK 序列,且命名為BdPAK。偵測其基因表現量和BdPAK螢光染色之結果顯示,BdPAK 之表現與位置與BdmGC-1 皆類似,表示BdPAK 與BdmGC-1可能有某種關聯。綜合上述結果可知,BdmGC-1 表現在翅、眼和足成蟲盤上並於蛹期第一天有較高表現,且可能與BdPAK 有某種關聯。zh_TW
dc.description.tableofcontents中文摘要 --- i ABSTRACT --- ii 目錄 ---iii 圖表目次 --- v 前言 ------------------------------------------------------------------------------- 1 前人研究 ------------------------------------------------------------------------- 2 一、 鳥苷酸環化酶之結構 ------------------------------------------ 2 二、 哺乳動物穿膜型GC --------------------------------------------- 2 三、 Rac/PAK/GC/cGMP ------------------------------------------- 3 四、 東方果實蠅鳥苷酸環化酶 ------------------------------------ 4 五、 成蟲盤之結構與分化 ------------------------------------------ 5 六、 成蟲盤分化之機制 --------------------------------------------- 6 七、 成蟲盤細胞增生調控 ------------------------------------------ 7 材料與方法 ---------------------------------------------------------------------- 8 一、 試驗蟲體 --------------------------------------------------------- 8 二、 RNA萃取 --------------------------------------------------------- 8 三、 反轉錄作用連鎖聚合酶反應 ------------------------------------- 9 四、 PAK簡併引子之設計 ---------------------------------------- 10 五、 cDNA膠體電泳------------------------------------------------- 10 六、 cDNA回收 ------------------------------------------------------ 10 七、 cDNA選殖與定序 ------------------------------------------- 11 八、 即時定量連鎖聚合酶反應 ----------------------------------- 12 九、 西方墨點法 ----------------------------------------------------- 13 十、 組織免疫螢光染色 -------------------------------------------- 14 結果 ----------------------------------------------------------------------------- 16 一、 翅、眼和足成蟲盤BdmGC-1 mRNA與蛋白之偵測 --------- 16 二、 翅、眼和足成蟲盤BdmGC-1於化蛹初期之表現 ------------- 16 三、 東方果實蠅BdPAK基因之選殖與表現 ------------------------ 17 四、 翅、眼和足成蟲盤之BdmGC-1和BdPAK的蛋白質分布 -------------------------------------------------------------------------- 17 討論 --------------------------------------------------------------------------- 19 一、 BdmGC-1於成蟲盤內的分布及各時期表現 ------------------ 19 二、 BdPAK於成蟲盤內的分布及各時期表現 --------------------- 20 參考文獻 ----------------------------------------------------------------------- 23 表 -------------------------------------------------------------------------------- 26 圖 -------------------------------------------------------------------------------- 28zh_TW
dc.subjectBactrocera dorsalisen_US
dc.subjectimaginal discen_US
dc.titleExpression of BdmGC-1 in three imaginal discs of the oriental fruit fly, Bactrocera dorsalis (Hendel)en_US
dc.typeThesis and Dissertationzh_TW
item.openairetypeThesis and Dissertation-
item.fulltextno fulltext-
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