基于超声波功率的大肠杆菌转化效率定量模型的开发与机理探索

IF 8.7 1区 化学 Q1 ACOUSTICS
Feifan Leng , Yubo Wang , Ning Zhu , Xiaopeng Guo , Wen Luo , Yonggang Wang
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引用次数: 0

摘要

超声波介导的质粒转化是一种具有广阔应用前景的微生物转化策略,已引起各个领域的兴趣。目前关于建立定量模型以了解转化效率与超声功率之间关系的研究还很有限。在不损伤质粒的超声波范围内,最大转化效率达到 4.84 × 105 CFU/μg DNA。利用基于膜渗透性变化的动力学模型确定了不同功率水平下的膜渗透性。结果表明,在特定范围内,超声功率、转化效率和膜渗透性之间呈线性相关。根据超声功率和大肠杆菌的转化效率建立了定量关系模型。电子显微镜显示,经过超声波处理的大肠杆菌细胞表现出孔隙形成和细胞扩张。此外,随着超声波功率的增加,细菌膜的完整性也会受到影响。在大肠杆菌 DH5α 中发现了九个与细胞膜成分和跨膜运输功能相关的基因。根据qRT-PCR结果,具有这些功能的基因(包括cusC、uidC、tolQ、tolA、ompC、yaiY)在超声波介导的大肠杆菌DH5α转化过程中起着关键作用。该研究表明,超声介导的大肠杆菌 DH5α 转化不是一个简单的物理化学过程,而是涉及反应膜相关基因的调控。这项研究为今后全面研究超声介导转化的分子机理奠定了基础,并为超声技术在基因工程及相关领域的应用提供了启示。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development and mechanism exploration of a quantitative model for Escherichia coli transformation efficiency based on ultrasonic power
Ultrasonic-mediated plasmid transformation is a promising microbial transformation strategy with broad application prospects that has attracted interest across various fields. Limited research exists on developing a quantitative model to understand the relationship between transformation efficiency and ultrasonic power. Within the ultrasonic range that did not damage plasmids, the maximum transformation efficiency reached at 4.84 × 105 CFU/μg DNA. A kinetic model based on changes in membrane permeability was utilized to determine the membrane permeability at different power levels. The results indicated a linear correlation between ultrasonic power, transformation efficiency, and membrane permeability within a specific range. A quantitative relationship model was established based on ultrasonic power and transformation efficiency in E. coli. Electron microscopy revealed that E. coli cells subjected to ultrasonic treatment exhibited pore formation and cellular expansion. Furthermore, the integrity of the bacterial membrane was compromised as ultrasonic power increased. Nine genes associated with the functional terms of cell membrane components and transmembrane transport were identified in E. coli DH5α. According to qRT-PCR results, genes with these functions (including cusC, uidC, tolQ, tolA, ompC, yaiY) play crucial roles in ultrasound-mediated transformation of E. coli DH5α. This study suggested that ultrasound-mediated transformation in E. coli DH5α is not a simple physical–chemical process but rather involves the regulation of responsive membrane-related genes. This research establishes the groundwork for future comprehensive investigations into the molecular mechanism of ultrasound-mediated transformation and provides insights for the application of ultrasound technology in genetic engineering and related fields.
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来源期刊
Ultrasonics Sonochemistry
Ultrasonics Sonochemistry 化学-化学综合
CiteScore
15.80
自引率
11.90%
发文量
361
审稿时长
59 days
期刊介绍: Ultrasonics Sonochemistry stands as a premier international journal dedicated to the publication of high-quality research articles primarily focusing on chemical reactions and reactors induced by ultrasonic waves, known as sonochemistry. Beyond chemical reactions, the journal also welcomes contributions related to cavitation-induced events and processing, including sonoluminescence, and the transformation of materials on chemical, physical, and biological levels. Since its inception in 1994, Ultrasonics Sonochemistry has consistently maintained a top ranking in the "Acoustics" category, reflecting its esteemed reputation in the field. The journal publishes exceptional papers covering various areas of ultrasonics and sonochemistry. Its contributions are highly regarded by both academia and industry stakeholders, demonstrating its relevance and impact in advancing research and innovation.
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