Ultrasound-mediated DNA transfer for bacteria
In environmental microbiology, the most commonly used methods of bacterial DNA transfer are conjugation and electroporation. However, conjugation requires physical contact and cell-pilus-cell interactions; electroporation requires low-ionic strength medium and high voltage. These limitations have ha...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
Oxford University Press
2007
|
| Subjects: |
| _version_ | 1797070618644971520 |
|---|---|
| author | Song, Y Hahn, T Thompson, I Mason, T Preston, G Li, G Paniwnyk, L Huang, W |
| author_facet | Song, Y Hahn, T Thompson, I Mason, T Preston, G Li, G Paniwnyk, L Huang, W |
| author_sort | Song, Y |
| collection | OXFORD |
| description | In environmental microbiology, the most commonly used methods of bacterial DNA transfer are conjugation and electroporation. However, conjugation requires physical contact and cell-pilus-cell interactions; electroporation requires low-ionic strength medium and high voltage. These limitations have hampered broad applications of bacterial DNA delivery. We have employed a standard low frequency 40 kHz ultrasound bath to successfully transfer plasmid pBBR1MCS2 into Pseudomonas putida UWC1, Escherichia coli DH5α and Pseudomonas fluorescens SBW25 with high efficiency. Under optimal conditions: ultrasound exposure time of 10 s, 50 mM CaCl2, temperature of 22°C, plasmid concentration of 0.8 ng/μl, P. putida UWC1 cell concentration of 2.5 x 10^9 CFU (colony forming unit)/nl and reaction volume of 500 μl, the efficiency of ultrasound DNA delivery (UDD) was 9.8 ± 2.3 x 10^-6 transformants per cell, which was nine times more efficient than conjugation, and even four times greater than electroporation. We have also transferred pBBR1MCS2 into E. coli DH5α and P. fluorescens SBW25 with efficiencies of 1.16 ± 0.13 x 10^-6 and 4.33 ± 0.78 x 10^-6 transformants per cell, respectively. Low frequency UDD can be readily scaled up, allowing for the application of UDD not only in laboratory conditions but also on an industrial scale. |
| first_indexed | 2024-03-06T22:41:29Z |
| format | Journal article |
| id | oxford-uuid:5bb5f0b6-3d90-49d3-b5d1-1c3680261fda |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:41:29Z |
| publishDate | 2007 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | oxford-uuid:5bb5f0b6-3d90-49d3-b5d1-1c3680261fda2022-03-26T17:23:43ZUltrasound-mediated DNA transfer for bacteriaJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:5bb5f0b6-3d90-49d3-b5d1-1c3680261fdaPlant SciencesLife SciencesEnglishOxford University Research Archive - ValetOxford University Press2007Song, YHahn, TThompson, IMason, TPreston, GLi, GPaniwnyk, LHuang, WIn environmental microbiology, the most commonly used methods of bacterial DNA transfer are conjugation and electroporation. However, conjugation requires physical contact and cell-pilus-cell interactions; electroporation requires low-ionic strength medium and high voltage. These limitations have hampered broad applications of bacterial DNA delivery. We have employed a standard low frequency 40 kHz ultrasound bath to successfully transfer plasmid pBBR1MCS2 into Pseudomonas putida UWC1, Escherichia coli DH5α and Pseudomonas fluorescens SBW25 with high efficiency. Under optimal conditions: ultrasound exposure time of 10 s, 50 mM CaCl2, temperature of 22°C, plasmid concentration of 0.8 ng/μl, P. putida UWC1 cell concentration of 2.5 x 10^9 CFU (colony forming unit)/nl and reaction volume of 500 μl, the efficiency of ultrasound DNA delivery (UDD) was 9.8 ± 2.3 x 10^-6 transformants per cell, which was nine times more efficient than conjugation, and even four times greater than electroporation. We have also transferred pBBR1MCS2 into E. coli DH5α and P. fluorescens SBW25 with efficiencies of 1.16 ± 0.13 x 10^-6 and 4.33 ± 0.78 x 10^-6 transformants per cell, respectively. Low frequency UDD can be readily scaled up, allowing for the application of UDD not only in laboratory conditions but also on an industrial scale. |
| spellingShingle | Plant Sciences Life Sciences Song, Y Hahn, T Thompson, I Mason, T Preston, G Li, G Paniwnyk, L Huang, W Ultrasound-mediated DNA transfer for bacteria |
| title | Ultrasound-mediated DNA transfer for bacteria |
| title_full | Ultrasound-mediated DNA transfer for bacteria |
| title_fullStr | Ultrasound-mediated DNA transfer for bacteria |
| title_full_unstemmed | Ultrasound-mediated DNA transfer for bacteria |
| title_short | Ultrasound-mediated DNA transfer for bacteria |
| title_sort | ultrasound mediated dna transfer for bacteria |
| topic | Plant Sciences Life Sciences |
| work_keys_str_mv | AT songy ultrasoundmediateddnatransferforbacteria AT hahnt ultrasoundmediateddnatransferforbacteria AT thompsoni ultrasoundmediateddnatransferforbacteria AT masont ultrasoundmediateddnatransferforbacteria AT prestong ultrasoundmediateddnatransferforbacteria AT lig ultrasoundmediateddnatransferforbacteria AT paniwnykl ultrasoundmediateddnatransferforbacteria AT huangw ultrasoundmediateddnatransferforbacteria |