Development and mechanism exploration of a quantitative model for Escherichia coli transformation efficiency based on ultrasonic power

作者：Feifan Leng, Yubo Wang, Yubo Wang, Ning Zhu, Xiao-Peng Guo, Wen Luo, Yonggang Wang, Yonggang Wang · 发表于：Ultrasonics Sonochemistry · 年份：2024 · DOI：10.1016/j.ultsonch.2024.107132 · 被引用次数：6 · 研究领域：Viral Infectious Diseases and Gene Expression in Insects、Microbial Inactivation Methods、thermodynamics and calorimetric analyses

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 5 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 phys...