2008-01-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/682345摘要：本計畫旨在選殖蚜蟲具致死潛力之基因，並發展出檢測基因功能之分析平台。我們選擇以豌豆蚜蟲 (Acyrthosiphon pisum) 計畫執行之標的昆蟲，因豌豆蚜蟲為大宗檢防疫害蟲之一，而且豌豆蚜蟲之序列標幟資料庫的基因資料較其他種蚜蟲完整。基於調控早期發育的基因突變在模式動物中已被證實會導致胚胎死亡，我們推測此現象亦可能發生於蚜蟲，因此選殖蚜蟲早期發育相關的基因為本計畫的首要目標。運用生物資訊的與比較基因體的策略，我們將由現存豌豆蚜之基因庫找出標的基因，再利用分子遺傳之技術將基因選殖出來。第一階段之分析我們將建立整體原位雜合法之技術平台並運用此它來標定基因表現之時期與位置。 第二階段之分析重點在於建立蚜蟲腹腔注射核酸干擾術之技術平台並分析基因是否具備致死功能。我們希望整合上述這兩種分析蚜蟲基因之技術平台，作為爾後檢測蚜蟲基因之重要工具。<br> Abstract: We aim to clone genes with lethal potential and establish technology platforms for functional assay in aphids. In this study, the pea aphid Acyrthosiphon pisum is selected as the target insect partly because it is one of the most popular pest insects and partly because the Expressed Sequence Tag (EST) database of A. pisum has the most abundant collection of gene sequences among all aphid species in the GenBank. According to the fact that the mutation of genes involved in early development leads to the death of model animals, we presume this phenomenon is also conserved in aphids and thus the cloning of early genes becomes the most important task of this project. Combining the strategies of bioinformatics and comparative genomics, we aim to retrieve homologous genes with the potential to regulate early development. Cloning of the target genes from A. pisum is achieved via molecular-genetics tools. In parallel, we will establish a technology platform of whole-mount in situ hybridization as to detect the temporal and spatial expression of genes. Furthermore, we propose to develop intra-abdominal RNA interference in A. pisum and utilize this technology to select genes with lethal functions. We expect that the integration of whole-mount in situ hybridization and RNA interference in A. pisum will become a powerful tool for analyzing gene functions in aphids.蚜蟲致死基因整體原位雜合法核酸干擾術aphidlethal genewhole-mount in situ hybridizationRNA interference