頁籤選單縮合
| 題 名 | 紅麴菌Monascus purpureus NTU 568電轉型基因操作平臺開發=Development of Monascus purpureus NTU 568 Gene Manipulation Platform Using Electroporation Method |
|---|---|
| 作 者 | 潘意淩; 林志輝; | 書刊名 | 臺東大學綠色科學學刊 |
| 卷 期 | 14:2 2024.11[民113.11] |
| 頁 次 | 頁53-67 |
| 分類號 | 369.36 |
| 關鍵詞 | 紅麴菌; 次級代謝物; 同源重組; 電轉型效率; Monascus; Secondary metabolites; Gene manipulation; Electroporation efficiency; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 絲狀真菌因其能合成多種對人類有益的蛋白質和次級代謝物,而成為高商業價值 的工業生產者。紅麴菌是亞洲發酵食品的重要菌種,其產品廣泛用於膳食補充品和食 品著色劑。隨著全基因體序列的揭露,紅麴菌的次級代謝物生合成基因研究逐漸深入, 為功能性基因研究奠定了基礎。然而過去紅麴菌基因的主流操作工具–農桿菌轉型法, 由於其隨機嵌入的特性,並不適合進行需要精確操作的功能基因體研究。因此在深入 探討紅麴菌基因機制之前,開發一個精確有效的基因操作平台是必要的。本研究構建 了同源重組剔除質體 pHRL,針對紅麴菌 NTU 568 中的 pksCT 基因進行同源重組剔 除,成功降低了 89%的 citrinin 產量。研究結果顯示,去除細胞壁、使用線型質體以及 電轉後的再生培養時間是 NTU568 電轉型效率的關鍵因素,未來可進一步探討替換啟 動子或更有效的篩選標記基因,以提高穩定轉型株的篩選效率。 |
| 英文摘要 | Filamentous fungi have become important industrial producers with high commercial value due to their ability to synthesize various beneficial proteins and secondary metabolites. Among them, Monascus is a key strain in Asian fermented foods, with its products widely used as dietary supplements and food coloring agents. As the whole genome sequence has been deciphered, research into the biosynthesis genes of secondary metabolites in Monascus has gradually deepened, laying the foundation for precise genetic studies. However, the current gene manipulation tool for Monascus - Agrobacterium transformation method has limitations due to its random insertion characteristics, making it unsuitable for functional genomic research that requires precision. Therefore, developing an effective gene manipulation platform is essential before delving into the genetic mechanisms of Monascus. This study constructed a homologous recombination deletion plasmid, pHRL, targeting the pksCT gene in Monascus NTU 568, successfully reducing citrinin yield by 89%. The results indicate that cell wall removal, the use of linear plasmids, and regeneration culture time postelectroporation are key factors influencing the electroporation efficiency of NTU 568. Future research may explore alternative promoters or more effective selection markers to enhance the screening efficiency of stable transformants. |
本系統中英文摘要資訊取自各篇刊載內容。