雷帕霉素基因簇的异源表达

论文编号:SW151  论文字数:7941,页数:26

 摘 要：Rapamycin是一种由吸水链霉菌(Str.hygroscopicus)产生的含氮的非典型三烯类大环内酯类抗生素，显示抗真菌、抗炎、抗肿瘤和免疫抑制性能。虽然这些小分子药物在抗肿瘤及免疫抑制等方面表现出极好的效果，[1]但是，它们的天然合成量普遍较低，因此药用成本极高，无法进行实际应用。
 通过应用基因簇转移技术，具体来讲是通过构建细菌人工染色体（BAC）文库，我们已分别将雷帕霉素(rapamycin，RAP)这种小分子药物的整个生物合成基因簇转移到几种模式链霉菌中（白色链霉菌S. albus，天蓝色链霉菌S. ceolicolor M512，变铅青链霉菌S. lividans K4-114），得到了包含他们整个生物合成基因簇的基因工程菌。S. albus 28-1-1已经表达出某种具有生物活性的物质，如果能将其分离鉴定，将是非常有意义的工作。
 
关键词：雷帕霉素；免疫抑制剂；异源表达；优化；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
 
目  录
中文摘要 I                  
英文摘要 II
目录 III
1. 绪论 1
 1.1 目的和意义 1
 1.2 雷帕霉素性质 1
 1.3 雷帕霉素合成途径 4
 1.4 转基因簇技术和组合生物合成 5
 1.5 培养条件的优化 6
 1.6 我们研究的基础 7
2. 实验部分 8
 2.1 实验材料 8
 2.2 培养条件 10
 2.3 分析方法 11
3. 结果与讨论 14
 3.1 优化前发酵液检测 14
 3.2 条件优化后结果 16
 3.3 质谱分析 17
4. 结论与展望 20
致谢 21
参考文献 21

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