通过将traJ纳入可调动等离子体中,增强大肠杆菌与Streptomyces coelicolor的结合
Paula Valdés-Chiara1, Yago Concha1, Sergio Alonso-Fernández1
1Department of Functional Biology, Microbiology Area, IUOPA and ISPA, Faculty of Medicine, Universidad de Oviedo, c/ Julian Claveria 6, 33006, Oviedo, Spain.
Applied microbiology and biotechnology
|December 9, 2025
概括
将traJ基因添加到可调动的等离子体中显著提高了Streptomyces结合中的DNA转移效率. 这种提高,高达100倍,有助于基因工程和自然产品的发现在这些重要的细菌.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 在生产生物活性天然产品方面,Streptomyces物种至关重要.
- 无效的DNA传递方法阻碍了Streptomyces的基因操纵.
- 来自大肠杆菌的等离子体结合是将DNA引入Streptomyces的一个关键方法.
研究的目的:
- 研究将traJ基因纳入可调动等离子体对Streptomyces结合效率的影响.
- 为了确定traJ是否增强了等离子体转移速率,并克服了大肠杆菌对Streptomyces结合的限制.
主要方法:
- 改造了含有ORT的等离子体 (pRASK-SP44,pHL734),以包括traJ基因.
- 在pCER向量中引入了traJ的第二个副本.
- 通过测量经过大肠杆菌转移到Streptomyces后的跨结合体数量来评估结合效率.
主要成果:
- 将traJ纳入pRASK-SP44和pHL734增加了转结合物数量,分别是10倍和100倍.
- 将traJ的第二个副本添加到pCER中,使得等离子体转移提高了5倍.
- traJ 的存在和拷贝数与结合效率呈正相关.
结论:
- 将traJ纳入可调动等离子体中显著提高了Streptomyces的结合效率.
- 调节traJ水平提供了一种简单的策略,以提高Streptomyces的遗传可访问性.
- 这些发现支持开发用于Streptomyces基因组工程和自然产品发现的改进基因工具.
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