臺灣博碩士論文加值系統

參考已發表之鳥苷酸環化酶基因保留序列設計一對簡併引子 (degenerate primer)，以東方果實蠅 (Bactrocera dorsalis (Hendel)) 頭部總量RNA進行反轉錄作用 (reverse transcription) 和聚合酵素鏈鎖反應 (polymerase chain reaction, PCR)，得到一水溶性鳥苷酸環化酶基因cDNA，稱為Bdsgc1 (accession number為AY324653)。Bdsgc1具有2680 bp，內含一完整之開放讀架 (opening reading frame)，所衍生出的蛋白質具676個胺基酸，分子量約為75.62 kDa。將Bdsgc1與黃果蠅 (Drosophila melanogaster)、菸草天蛾 (Manduca sexta)、人類(Homo sapiens) 及老鼠 (Rattus norvegicus) 之水溶性鳥苷酸環化酶胺基酸序列比對，結果顯示Bdsgc1和上述之鳥苷酸環化酶胺基酸序列之相似度及一致性皆為58%以上，因此更可確認所得之Bdsgc1應為東方果實蠅之鳥苷酸環化酶基因。
以Bdsgc1 5’端0.9 kb大小之核酸片段為探針，利用北方墨點法 (Northern blot) 偵測Bdsgc1在東方果實蠅蟲體內之表現，結果顯示Bdsgc1僅在成蟲期表現，卵、幼蟲及蛹期皆無表現。成蟲體內又以頭部為其主要表現之區域，胸部及腹部皆無偵測到該基因之表現。利用相同之核酸片段作為探針進行原位雜合 (in situ hybridization) 試驗，觀察Bdsgc1在東方果實蠅頭部內之分布情形，結果顯示Bdsgc1 之mRNA專一性表現於腦區周圍的神經細胞本體，尤以視葉及嗅葉最為顯著。

A cDNA clone encoding for a soluble guanylyl cyclase was isolated from the head of Oriental fruit fly, Bactrocera dorsalis (Hendel), named Bdsgc1 (accession number is AY324653). Bdsgc1 consits of 2680 bp, which contains an open reading frame and encodes a protein of 676 amino acids with a predicted molecular weight of 75.62 kDa. Both identity and similarity of Bdsgc1 are greater than 58% in comparison to Drosophila melanogaster, Rattus norvegicus, Homo sapiens and Manduca sexta soluble guanylyl cyclases, suggesting that the Bdsgc1 is an Oriental fruit fly soluble guanylyl cyclase.
The results of northern blot demonstrated that the Bdsgc1 mRNA was expressed in the head of adult B. dorsalis. It was absent in the eggs, larvae and pupa. In the adult stage, there was no significant difference between males and females. In situ hybridization analyses revealed that Bdsgc1 mRNA mainly expressed in the nerve cell bodies of the brain of adult B. dorsalis, especially, in the medulla, lobula and antennal lobe. The expression pattern may indicate that the Bdsgc1 involves in visual and olfactory signal transmission.

目錄
中文摘要 -----------------------------------------------------1
英文摘要 -----------------------------------------------------2
前言 ---------------------------------------------------------3
前人研究 -----------------------------------------------------5
材料與方法 ------------------------------------------------- 10
結果 --------------------------------------------------------24
討論 --------------------------------------------------------29
參考文獻 ----------------------------------------------------32
圖表 --------------------------------------------------------38
附錄 --------------------------------------------------------52

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