The effect of plasmid DNA sizes and other factors on electrotransformation of Escherichia coli JM109

Szostková, Monika; Horáková, Danuška

doi:10.1016/s0302-4598(98)00203-7

Cited by 23 publications

(15 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The corresponding transformation efficiency was 4660±341 transformants/μg DNA. We also found that the electroporation with the electric field strength of more than 14 kV/cm was apt to cause a number of complications, such as arcing and shortening of the electrical pulses, as reported by Szostková and Horáková .…”

Section: Resultssupporting

confidence: 78%

Enhanced electrotransformation of the ethanologen Zymomonas mobilis ZM4 with plasmids

Zou

Zhang

You

et al. 2012

Engineering in Life Sciences

View full text Add to dashboard Cite

Enhanced electrotransformation of the ethanologen Zymomonas mobilis ZM4 with plasmidsThe Zymomonas mobilis ZM4 strain with excellent ethanol-producing capabilities was the first strain of Z. mobilis, which was sequenced. This strain is resistant to transformation, and no previous study has shown a detailed protocol for electrotransfer of ZM4 with foreign DNA. In this work, many electrical and biological parameters were selected and evaluated in order to optimize the electrotransformation of ZM4. First, improved transformation efficiencies of 11 896, 99, 96 and 5989 transformants/mg DNA were separately achieved with shuttle plasmid pZB21-mini (3082 bp), pZB21 (5930 bp), pZA22 (6994 bp) and broad-host-range vector pBBR1MCS-2 (5144 bp) all prepared from Escherichia coli JM110. The crucial factors affecting the transformation efficiency included the source of the plasmid (the best strain was ZM4), origin and size of the plasmids, growth phase of the cells (the most ideal phase was early log phase with OD 600 of 0.3-0.4), the electric field strength (generally 11.75 kV/cm-13.25 kV/cm) and the recovery time (3-24 h). Further, based upon the optimal transformation protocol mentioned above for replicative plasmids in ZM4, (i) the electrotransformation by recombinant plasmid pBBR1MCS-2-Pgap-FLP (6880 bp) was an immediate success with the transformation efficiency 10 2 transformants/mg DNA; (ii) the site-specific integration efficiencies (expressed in terms of ''per mg of DNA'') of 3-6 integrating transformants was obtained using the integrating plasmid pBR328-ldhR-cml-ldhL (7447 bp). This study will assist genetic and biotechnological research of ZM4 and other Z. mobilis strains by providing information about suitable vectors and a more universal and reliable procedure for introducing DNA into this strain.

show abstract

Section: Resultssupporting

confidence: 78%

Enhanced electrotransformation of the ethanologen Zymomonas mobilis ZM4 with plasmids

Zou

Zhang

You

et al. 2012

Engineering in Life Sciences

View full text Add to dashboard Cite

show abstract

“…This prompted us to develop a functional conjugative system to introduce the vector into bifidobacteria, as conjugation has at least two important advantages over electroporation. First, electroporation efficiencies are known to decrease as the size of the vector increases (Szostková & Horakova, 1998). Conjugation does not have this limitation and has been successfully used to transfer entire genomes in E. coli (Isaacs et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Developing an efficient and reproducible conjugation-based gene transfer system for bifidobacteria

Domínguez

O’Sullivan

2013

Microbiology

View full text Add to dashboard Cite

“…Two classical expressions of the transformation are used (Szostkova and Horakova, 1998). The efficiency of transformation is reflected by the number of transformants I Jlg of used DNA and the transformation frequency is calculated by the percentage of survival cells which have been transformed.…”

Section: Expression Of Resultsmentioning

confidence: 99%

“…Effect is observed in the total response in terms of number of cells electrotransformed. Many experiments appear to show that the transformation efficiency decreased with an increase in the size ofDNA, but a limit to this conclusion is that variation of the size is obtained by use of different plasmids with various antibiotic markers and origins (McLaughlin andFerretti, 1995, Szostkova andHorakova, 1998). This observation is not obvious when different plasmid sizes were obtained either by ligation of the same marker region in various plasmids or by successive deletion in a given plasmid (Allen andBlaschek, 1990, Ohse, et al, 1995).…”

Section: Effects Of Size and Molecular Form Of Dna And Plasmid Concenmentioning

confidence: 99%

General Principles of Bacteria Electrotransformation: Key Steps

Eynard

Teissié

2000

Electrotransformation of Bacteria

View full text Add to dashboard Cite

The effect of plasmid DNA sizes and other factors on electrotransformation of Escherichia coli JM109

Cited by 23 publications

References 17 publications

Enhanced electrotransformation of the ethanologen Zymomonas mobilis ZM4 with plasmids

Enhanced electrotransformation of the ethanologen Zymomonas mobilis ZM4 with plasmids

Developing an efficient and reproducible conjugation-based gene transfer system for bifidobacteria

General Principles of Bacteria Electrotransformation: Key Steps

Contact Info

Product

Resources

About