雷帕霉素基因簇的異源表達

生物工程論文編號:SW151  論文字數:7941,頁數:26

目  錄
中文摘要 I                  
英文摘要 II
目錄 III
1. 緒論 1
 1.1 目的和意義 1
 1.2 雷帕霉素性質 1
 1.3 雷帕霉素合成途徑 4
 1.4 轉基因簇技術和組合生物合成 5
 1.5 培養條件的優化 6
 1.6 我們研究的基礎 7
2. 實驗部分 8
 2.1 實驗材料 8
 2.2 培養條件 10
 2.3 分析方法 11
3. 結果與討論 14
 3.1 優化前發酵液檢測 14
 3.2 條件優化后結果 16
 3.3 質譜分析 17
4. 結論與展望 20
致謝 21
參考文獻 21

 摘 要：Rapamycin是一種由吸水鏈霉菌(Str.hygroscopicus)產生的含氮的非典型三烯類大環內酯類抗生素，顯示抗真菌、抗炎、抗腫瘤和免疫抑制性能。雖然這些小分子藥物在抗腫瘤及免疫抑制等方面表現出極好的效果，[1]但是，它們的天然合成量普遍較低，因此藥用成本極高，無法進行實際應用。
 通過應用基因簇轉移技術，具體來講是通過構建細菌人工染色體（BAC）文庫，我們已分別將雷帕霉素(rapamycin，RAP)這種小分子藥物的整個生物合成基因簇轉移到幾種模式鏈霉菌中（白色鏈霉菌S. albus，天藍色鏈霉菌S. ceolicolor M512，變鉛青鏈霉菌S. lividans K4-114），得到了包含他們整個生物合成基因簇的基因工程菌。S. albus 28-1-1已經表達出某種具有生物活性的物質，如果能將其分離鑒定，將是非常有意義的工作。
 
關鍵詞：雷帕霉素；免疫抑制劑；異源表達；優化；基因簇
 
Heterologous expression of rapamycin gene cluster
 
Abstract: Rapamycin, a macrolide antibiotic produced by Streptomyces hygroscopicus, was initially discovered as an antifungal agent and subsequently shown to exhibit potent antitumor and immunosuppressive activities. Although these small molecule drugs in the anti-tumor and immune suppression, and so have shown excellent results, but their natural synthesis is generally low, high medical costs, not for practical applications.
 Through the application of gene cluster transfer technology, specifically, through the construction of bacterial artificial chromosome (BAC) library, we have respectively rapamycin (rapamycin, RAP) this small molecule drug synthesis of the entire gene cluster transferred to several models Streptomyces (white Streptomyces S. albus, Blue Streptomyces S. ceolicolor M512, variable lead Green Streptomyces S. lividans K4-114), has been with their biological synthesis of the entire gene cluster of genetic engineering bacteria.We expect the better results.
Keywords：Rapamycin  immunosuppressant  heterologous expression  optimization  gene cluster