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使用高通量诱导屏幕从Streptomyces albus J1074中发现一种神秘的抗真菌化合物
Fei Xu1, Behnam Nazari1, Kyuho Moon1
1Department of Chemistry, Princeton University , Princeton, New Jersey 08544, United States.
Journal of the American Chemical Society
|June 8, 2017
概括
研究人员开发了一种高通量选方法 (HiTES),以激活Streptomyces细菌中的静态基因集群. 这种方法通过发现特定路径的抑制剂来确定新的天然产品,包括抗真菌药物和向癌症相关蛋白酶的化合物.
科学领域:
- 微生物学
- 自然产品的发现
- 合成生物学
背景情况:
- 许多微生物生物合成基因集群在实验室条件下保持沉默,限制了新型自然产品的发现.
- 激活这些沉默的基因集群对于发现新的生物活性化合物至关重要.
- 之前的高通量查 (HiTES) 方法对格拉姆阴性细菌有效,但尚未应用于Streptomyces.
研究的目的:
- 在Streptomyces中开发和实施高通量转录试验,以激活静态基因群.
- 利用HiTES方法发现小分子诱导物和新型二次代谢物.
- 调查静态基因集群激活的机制,并确定调控因素.
主要方法:
- 开发了使用EGFP作为二次代谢物合成的高通量转录试验.
- 对Streptomyces albus J1074进行化学基因高通量选 (HiTES) 方法.
- 通过激活静态基因集群产生的新型小分子的鉴定和特征.
主要成果:
- 使用HiTES方法成功激活了Streptomyces albus J1074中的一个静态基因集群.
- 埃托波西德和艾弗梅克丁被确定为二次代谢产生的强有力的诱导剂.
- 隔离和结构特征14个新型小分子,包括一种新的抗真菌和囊蛋白酶抑制剂.
结论:
- 开发的HiTES试验有效地激活了Streptomyces中的静态基因集群.
- 这种方法有助于发现具有潜在药物应用的新型天然产品.
- 鉴定了一种通路特异性的转录抑制剂,负责沉默基因,为基因调控提供了洞察力.
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