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研究生:陳仕錤
研究生(外文):Shyh-Chi Chen
論文名稱:甲殼類生物胺受體之分子選殖
論文名稱(外文):Molecular Cloning of Crustacean Biogenic Amine Receptors
指導教授:李奇英
指導教授(外文):Chi-Ying Lee
學位類別:碩士
校院名稱:國立彰化師範大學
系所名稱:生物學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:67
中文關鍵詞:生物胺生物胺受體G蛋白偶合受體血清素多巴胺章魚涎胺甲殼類克式螯蝦
外文關鍵詞:biogenic aminebiogenic amineG protein-coupled receptorserotonin5-HTdopamineoctopaminecrustaceanProcambarus clarkii
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生物胺是普遍存在於生物體中,調節多種生理功能之神經傳遞物質(neurotransmitter)。甲殼類動物眼柄中的一些生物胺,如多巴胺、血清素、章魚涎胺等,已被證實具有調節神經內分泌組織(X-器官-血竇腺)釋放太類荷爾蒙的功能。
本論文利用分子選殖方法,尋找眼柄神經節中的生物胺受體,以期更進一步釐清生物胺與X-器官-血竇腺之間活化、分泌及偶合的調控機制。藉由生物胺受體在第二、六、七個跨模區域(transmembrane domain)的保守性序列,我們設計了用於聚合連鎖反應(PCR)的引子(primer)。PCR產物經選殖、純化、定序之後,發現它們的序列具有生物胺受體基因的特徵。根據以上這些序列,再進一步設計用以進行RACE(3’-and 5’-rapid amplification of cDNA ends)反應的引子。RACE PCR的產物經序列分析後發現與嗅覺受體(olfactory receptor)基因相似。這些所選殖到具生物胺受體基因特徵的序列片段,將用於篩選甲殼類眼柄的cDNA library,以期能獲得甲殼類生物胺受體基因的全長序列。

Biogenic amines, functioning as eurotransmitters/neurohormones, are involved in a wide variety of physiological processes. In crustaceans, several biogenic amines, including serotonin (5-HT), dopamine, and octopamine have been found to regulate the release of several peptide hormones from a neuroendocrine tissues, X-organ-sinus gland complex, located within the eyestalks of decapods.
In order to advance our knowledge regarding neuronal regulation of hormonal secretion in crustaceans, the objective of this study was to molecularly clone biogenic amine receptors from the eyestalk ganglia, where the X-organ-sinus gland receives neuronal signals that subsequently regulate the secretion of neuropeptides. Primers were designed based on the conserved sequences located in the 2nd, 6th, 7th transmembrane domains of the biogenic amine receptors and used for PCR amplification of eyestalk ganglia cDNA. PCR products of expected size were purified, cloned, and sequenced. Sequence analysis revealed that they contain sequence characteristics of biogenic amine receptor genes, including octopamine/tyramine, dopamine, serotonin, and histamine receptors. Primers were further designed based on some of the amplified sequences and used for RACE amplification of 3’- and 5’-ends. However, sequence analysis of these RACE PCR products indicated that they share high similarity with olfactory receptors, but not biogenic amine receptors. Currently, the amplified sequences showing characteristics of biogenic amine receptors are used as probes in screening a cDNA library in order to obtain full-length sequences of novel crustacean biogenic amine receptor genes.

中文摘要------------------------------------------------1
Abstract -----------------------------------------------2
Introduction -------------------------------------------4
Materials and Methods ----------------------------------10
Results ------------------------------------------------16
Discussion ---------------------------------------------20
References ---------------------------------------------25
Table --------------------------------------------------36
Figure --------------------------------------------------44

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