Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells
ABSTRACT The transformation of DNA into cells is the basis of molecular biology. Commonly employed techniques include heat shock transformation, electro-transformation, conjugation, transduction, and protoplast fusion. Recently, ultrasonic transformation technology has been developed to transfer DNA...
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| Language: | English |
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American Society for Microbiology
2024-12-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.00978-24 |
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| author | Meng Zhang Rongkang Tang Fang-Xia Li Wen-Yu Jin Jia-Xin Guo Lin-Zuo Teng Guangxun Meng Philippe J. Sansonetti Yi-Zhou Gao |
| author_facet | Meng Zhang Rongkang Tang Fang-Xia Li Wen-Yu Jin Jia-Xin Guo Lin-Zuo Teng Guangxun Meng Philippe J. Sansonetti Yi-Zhou Gao |
| author_sort | Meng Zhang |
| collection | DOAJ |
| description | ABSTRACT The transformation of DNA into cells is the basis of molecular biology. Commonly employed techniques include heat shock transformation, electro-transformation, conjugation, transduction, and protoplast fusion. Recently, ultrasonic transformation technology has been developed to transfer DNA into competent cells. The transformation conditions, such as temperature and ultrasonic power, were preliminarily studied. However, this technique has not been widely applied because competent cells must be prepared de novo. In this study, various factors, such as ultrasonic frequency and power, were optimized for the ultrasonic transformation of Escherichia coli. The study found that the optimal conditions for ultrasonic transformation with a defined ultrasonic transformation vial were a frequency of 28 kHz and a power of 80 W. Meanwhile, this research demonstrated that combining the 42°C heat shock conditions with ultrasonic transformation is the most efficient method compared to using only heat shock. Furthermore, the cryoprotective agent ratio for ultrasonic competent cells was investigated and optimized. These findings provide new insights into enhancing transformation efficiency and lay a foundation for the broader application of ultrasonic transformation.IMPORTANCEPlasmid transformation is widely applicable in gene expression and modification. As an efficient, non-invasive, and gentle method of transformation, ultrasonic transformation provides a novel approach for strain modification. This research presents new strategies for enhancing transformation efficiency and lays the groundwork for expanding the utilization of ultrasonic transformation. |
| format | Article |
| id | doaj-art-a7929af282bf45af88bcc7cfee0c10a2 |
| institution | Kabale University |
| issn | 2165-0497 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-a7929af282bf45af88bcc7cfee0c10a22024-12-05T14:01:22ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972024-12-01121210.1128/spectrum.00978-24Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cellsMeng Zhang0Rongkang Tang1Fang-Xia Li2Wen-Yu Jin3Jia-Xin Guo4Lin-Zuo Teng5Guangxun Meng6Philippe J. Sansonetti7Yi-Zhou Gao8Shanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaShanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, ChinaABSTRACT The transformation of DNA into cells is the basis of molecular biology. Commonly employed techniques include heat shock transformation, electro-transformation, conjugation, transduction, and protoplast fusion. Recently, ultrasonic transformation technology has been developed to transfer DNA into competent cells. The transformation conditions, such as temperature and ultrasonic power, were preliminarily studied. However, this technique has not been widely applied because competent cells must be prepared de novo. In this study, various factors, such as ultrasonic frequency and power, were optimized for the ultrasonic transformation of Escherichia coli. The study found that the optimal conditions for ultrasonic transformation with a defined ultrasonic transformation vial were a frequency of 28 kHz and a power of 80 W. Meanwhile, this research demonstrated that combining the 42°C heat shock conditions with ultrasonic transformation is the most efficient method compared to using only heat shock. Furthermore, the cryoprotective agent ratio for ultrasonic competent cells was investigated and optimized. These findings provide new insights into enhancing transformation efficiency and lay a foundation for the broader application of ultrasonic transformation.IMPORTANCEPlasmid transformation is widely applicable in gene expression and modification. As an efficient, non-invasive, and gentle method of transformation, ultrasonic transformation provides a novel approach for strain modification. This research presents new strategies for enhancing transformation efficiency and lays the groundwork for expanding the utilization of ultrasonic transformation.https://journals.asm.org/doi/10.1128/spectrum.00978-24ultrasonic plasmid transformationultrasonic transformation containercryopreservationcryoprotective agents |
| spellingShingle | Meng Zhang Rongkang Tang Fang-Xia Li Wen-Yu Jin Jia-Xin Guo Lin-Zuo Teng Guangxun Meng Philippe J. Sansonetti Yi-Zhou Gao Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells Microbiology Spectrum ultrasonic plasmid transformation ultrasonic transformation container cryopreservation cryoprotective agents |
| title | Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells |
| title_full | Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells |
| title_fullStr | Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells |
| title_full_unstemmed | Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells |
| title_short | Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells |
| title_sort | optimization of ultrasound mediated dna transfer for bacteria and preservation of frozen competent cells |
| topic | ultrasonic plasmid transformation ultrasonic transformation container cryopreservation cryoprotective agents |
| url | https://journals.asm.org/doi/10.1128/spectrum.00978-24 |
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