請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/30685
標題: cDNA cloning and expression of the vitellogenin receptor from the oriental fruit fly, Bactrocera dorsalis
東方果實蠅之卵黃原蛋白受器cDNA序列分析及表現情形探討
作者: Lin, Wei-Jen
林威仁
關鍵字: Bactrocera dorsalis
東方果實蠅
vitellogenin receptor
ovary
fat body
卵黃原蛋白受器
卵巢
脂肪體
出版社: 昆蟲學系所
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摘要: 卵黃原蛋白透過受器媒介內吞作用吸收到卵內儲存供胚胎發育。本研究選殖東方果實蠅 (Bactrocera dorsalis Hendel) 卵黃原蛋白受器cDNA 序列並分析其表現情形。利用RACE得到一全長6585 bp之cDNA,其開放讀架 (open reading frame, ORF) 可轉譯出1923個胺基酸,其功能區域排列符合昆蟲卵黃原蛋白受器之特性,與黃果蠅 (Drosophila melanogaster) 有極高之相似度 (70%)。利用RT-PCR分析,脂肪體和卵巢兩者皆會表現卵黃原蛋白受器mRNA。進一步以西方墨點法分析,顯示卵巢與脂肪體皆會表現卵黃原蛋白受器之蛋白質 (約213 kDa)。利用qRT-PCR分析於不同日齡 (羽化後十天內) 雌成蟲脂肪體內卵黃原蛋白受器mRNA之表現,結果顯示羽化後至六日齡會隨時間而增加,之後則會隨時間而下降;蛋白質表現則為羽化初期 (羽化後第零天到第二天) 表現量較低,第三天開始增加直到第六天有高量表現,隨後表現量開始下降。qRT-PCR分析於不同日齡 (羽化後十天內) 之雌成蟲卵巢內卵黃原蛋白受器mRNA之表現結果顯示,雌成蟲羽化後有高量表現 (二十四小時內) 之後持續下降直到第二天,第三天開始有上升之趨勢直到第十天。蛋白質的表現則為羽化初期之蛋白表現量較低,從第三天開始有上升的趨勢直到第十天。本研究結果指出卵黃原蛋白受器於卵巢發育時期會持續的表現,而其於脂肪體之功能可能為吸收血體液中之卵黃原蛋白。
The vitellogenin (Vg) is absorbed into the growing oocytes via receptor-mediated endocytosis for the embryonic development. In this study, vitellogenin receptor cDNA of the oriental fruit fly (Bactrocera dorsalis Hendel) was cloned and its expression was analyzed. Using RACE technology, a putative B. dorsalis vitellogenin receptor (BdVgR) was cloned. The BdVgR cDNA has a length of 6585 bp encoding 1923 amino acids. The BdVgR has conserved motif arrangement with other insect VgRs, and showed the high similarity to Drosophila VgR (70%). The PCR analysis showed that the mRNA of BdVgR was expressed in both ovary and fat body. The western blot analysis demonstrated that the BdVgR (~213 kDa) was expressed in both ovary and fat body. The qRT-PCR analysis showed that the expression of BdVgR in the fat body was low after eclosion.The BdVgR mRNA increased from day 4, and it showed the highest level on day 6, and then it decreased with time. The expression level of BdVgR in the ovary was high in 24h after eclosion, and then it decreased until day 2. It increased with time on day 3. The BdVgR protein was expressing highly in the fat body between day3 and day 6; but it gradually increased from day 3 in the ovary. In conclusions, the BdVgR was expressed during the ovary development. The function of BdVgR in the fat body could be involved in reuptake vitellogenins from the hemolymph.
URI: http://hdl.handle.net/11455/30685
其他識別: U0005-1208201102433300
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1208201102433300
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