Optimization and mechanism exploration for Escherichia coli transformed with plasmid pUC19 by the combination with ultrasound treatment and chemical method

Yonggang Wang; Shangchen Sun; Linmiao Yu; Shu Hu; Wenguang Fan; Feifan Leng; Jianzhong Ma

doi:10.1016/j.ultsonch.2021.105552

Optimization and mechanism exploration for Escherichia coli transformed with plasmid pUC19 by the combination with ultrasound treatment and chemical method

Ultrason Sonochem. 2021 Jun:74:105552. doi: 10.1016/j.ultsonch.2021.105552. Epub 2021 Apr 20.

Authors

Yonggang Wang¹, Shangchen Sun², Linmiao Yu³, Shu Hu³, Wenguang Fan³, Feifan Leng⁴, Jianzhong Ma⁵

Affiliations

¹ School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China. Electronic address: wangyg@lut.cn.
² School of Petrochemical Engineering, Lanzhou University of Technology, Lanzhou 730050, China.
³ School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China.
⁴ School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China. Electronic address: lff0978@sina.com.
⁵ School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China. Electronic address: majz@lut.cn.

Abstract

As a basic technique of molecular cloning, bio-transformation has been successfully used in the fields of biomedicine and food processing. In this study, we established a transformation system of exogenous DNA into E. coli cells mediated by ultrasound. Under the optimal conditions (i.e. 35 °C, 40 W, 25 s, OD600 = 0.4-0.6) optimized by RSM, the transformation efficiency reached at 1.006 × 10⁷ CFU/μg DNA. The results of membrane permeability, macromolecular substance and cell structure analysis before and after ultrasound treatment showed that the damage of host cells induced by lower (40 W) ultrasound and shorter ultrasound time (25 s) was reversible, and the transformation efficiency and cell survival rate were not significantly affected under this condition. In brief, proper changes in cell membrane and cell wall were the basic conditions for host cells to uptake exogenous DNA, while, whether exogenous DNA could be replicated and expressed in cells depends on the viability of host cells.

Keywords: Escherichia coli; Genetic transformation; Plasmid pUC19; Ultrasound treatment.

MeSH terms

Biotransformation
Escherichia coli / genetics*
Escherichia coli / metabolism*
Plasmids / genetics*
Ultrasonic Waves*