Optimization and mechanism exploration for Escherichia coli transformed with plasmid pUC19 by the combination with ultrasound treatment and chemical method
| Autor: | Shu Hu, Shangchen Sun, Jianzhong Ma, Linmiao Yu, Wenguang Fan, Yonggang Wang, Feifan Leng |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Acoustics and Ultrasonics
Membrane permeability QC221-246 Ultrasound treatment 02 engineering and technology Molecular cloning 010402 general chemistry medicine.disease_cause 01 natural sciences Inorganic Chemistry Genetic transformation chemistry.chemical_compound Plasmid medicine Escherichia coli Chemical Engineering (miscellaneous) Environmental Chemistry Radiology Nuclear Medicine and imaging Original Research Article QD1-999 Biotransformation ComputingMethodologies_COMPUTERGRAPHICS Organic Chemistry Acoustics. Sound 021001 nanoscience & nanotechnology 0104 chemical sciences Transformation (genetics) Chemistry chemistry Ultrasonic Waves Plasmid pUC19 Biophysics Exogenous DNA 0210 nano-technology DNA Transformation efficiency Plasmids |
| Zdroj: | Ultrasonics Sonochemistry Ultrasonics Sonochemistry, Vol 74, Iss, Pp 105552-(2021) |
| ISSN: | 1873-2828 1350-4177 |
| Popis: | Graphical abstract Highlights • The transfer method was optimized in ultrasound-mediated. • E. coli DH5α as the receptor, pUC19 as exogenous DNA. • The E. coli DH5α membrane permeability changed allowing exogenous DNA to entered the cytoplasm. 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. |
| Databáze: | OpenAIRE |
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