School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China.
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China; School of Petrochemical Engineering, Lanzhou University of Technology, Lanzhou 730050, China.
Ultrason Sonochem. 2024 Dec;111:107132. doi: 10.1016/j.ultsonch.2024.107132. Epub 2024 Oct 28.
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 × 10 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.
超声介导的质粒转化是一种有前途的微生物转化策略,具有广泛的应用前景,引起了各个领域的兴趣。目前关于开发一种定量模型来理解转化效率与超声功率之间关系的研究还很有限。在不破坏质粒的超声范围内,最大转化效率达到 4.84×10 CFU/μg DNA。利用基于膜通透性变化的动力学模型,确定了不同功率水平下的膜通透性。结果表明,在特定范围内,超声功率、转化效率和膜通透性之间存在线性相关。根据超声功率和大肠杆菌中的转化效率,建立了一个定量关系模型。电子显微镜显示,经过超声处理的大肠杆菌细胞形成孔并扩张。此外,随着超声功率的增加,细菌膜的完整性受到损害。在大肠杆菌 DH5α 中鉴定出与细胞膜成分和跨膜转运功能术语相关的 9 个基因。根据 qRT-PCR 结果,具有这些功能的基因(包括 cusC、uidC、tolQ、tolA、ompC、yaiY)在大肠杆菌 DH5α 的超声介导转化中发挥着关键作用。本研究表明,大肠杆菌 DH5α 的超声介导转化不是一个简单的物理化学过程,而是涉及响应性膜相关基因的调节。这项研究为进一步研究超声介导转化的分子机制奠定了基础,并为超声技术在基因工程和相关领域的应用提供了新的思路。