Nucleic Acids Research, 1988, Vol. 16, No. 13 6127-6145
© 1988
Articles |
High efficiency transformation of E.coli by high voltage electroporation
Molecular Biology Group, Bio-Rad Laboratories 1414 Harbour Way So., Richmond, CA 94804, USA 1Electrophoresis Engineering Group, Bio-Rad Laboratories 1414 Harbour Way So., Richmond, CA 94804, USA 2Department of Medical Microbiology, Stanford Medical center Stanford, CA 94305, USA
* To whom correspondence should be addressed
Received April 13, 1988. Revised May 31, 1988. Accepted May 31, 1988.
E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA to brief but intense electrical fields of exponential decay waveform (electroporation). We have obtained 109 to 1010 transformants/µg with strains LE392 and DH5
, and plasmids pUC18 and pBR329. The process is highly dependent on two characteristics of the electrical pulse: the electric field strength and the pulse length (RC time constant). The frequency of transformation is a linear function of the DNA concentration over at least six orders of magnitude; and the efficiency of transformation is a function of the cell concentration. Most of the surviving cells are competent with up to 80% transformed at high DNA concentration. The mechanism does not appear to include binding of the DNA to the cells prior to entry. Possible mechanisms are discussed and a simple procedure for the practical use of this technique is presented.
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D. O. d. O. Lucana, P. Zou, M. Nierhaus, and H. Schrempf Identification of a novel two-component system SenS/SenR modulating the production of the catalase-peroxidase CpeB and the haem-binding protein HbpS in Streptomyces reticuli Microbiology, November 1, 2005; 151(11): 3603 - 3614. [Abstract] [Full Text] [PDF]
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Z. Chen and H. Zhao A highly sensitive selection method for directed evolution of homing endonucleases Nucleic Acids Res., October 6, 2005; 33(18): e154 - e154. [Abstract] [Full Text] [PDF]
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P. Goffin, M. Deghorain, J.-L. Mainardi, I. Tytgat, M.-C. Champomier-Verges, M. Kleerebezem, and P. Hols Lactate Racemization as a Rescue Pathway for Supplying D-Lactate to the Cell Wall Biosynthesis Machinery in Lactobacillus plantarum J. Bacteriol., October 1, 2005; 187(19): 6750 - 6761. [Abstract] [Full Text] [PDF]
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M. E. Schreiner, D. Fiur, J. Holatko, M. Patek, and B. J. Eikmanns E1 Enzyme of the Pyruvate Dehydrogenase Complex in Corynebacterium glutamicum: Molecular Analysis of the Gene and Phylogenetic Aspects J. Bacteriol., September 1, 2005; 187(17): 6005 - 6018. [Abstract] [Full Text] [PDF]
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A. G. Torres, R. C. Vazquez-Juarez, C. B. Tutt, and J. G. Garcia-Gallegos Pathoadaptive Mutation That Mediates Adherence of Shiga Toxin-Producing Escherichia coli O111 Infect. Immun., August 1, 2005; 73(8): 4766 - 4776. [Abstract] [Full Text] [PDF]
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M. J. Volles and P. T. Lansbury Jr A computer program for the estimation of protein and nucleic acid sequence diversity in random point mutagenesis libraries Nucleic Acids Res., June 29, 2005; 33(11): 3667 - 3677. [Abstract] [Full Text] [PDF]
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P. Joseph, M. Ratnayake-Lecamwasam, and A. L. Sonenshein A Region of Bacillus subtilis CodY Protein Required for Interaction with DNA J. Bacteriol., June 15, 2005; 187(12): 4127 - 4139. [Abstract] [Full Text] [PDF]
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V. L. Taylor, R. W. Titball, and P. C. F. Oyston Oral immunization with a dam mutant of Yersinia pseudotuberculosis protects against plague Microbiology, June 1, 2005; 151(6): 1919 - 1926. [Abstract] [Full Text] [PDF]
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J. B. van Beilen, R. Holtackers, D. Luscher, U. Bauer, B. Witholt, and W. A. Duetz Biocatalytic Production of Perillyl Alcohol from Limonene by Using a Novel Mycobacterium sp. Cytochrome P450 Alkane Hydroxylase Expressed in Pseudomonas putida Appl. Envir. Microbiol., April 1, 2005; 71(4): 1737 - 1744. [Abstract] [Full Text] [PDF]
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P. Wunsch and W. G. Zumft Functional Domains of NosR, a Novel Transmembrane Iron-Sulfur Flavoprotein Necessary for Nitrous Oxide Respiration J. Bacteriol., March 15, 2005; 187(6): 1992 - 2001. [Abstract] [Full Text] [PDF]
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R. van Kranenburg, N. Golic, R. Bongers, R. J. Leer, W. M. de Vos, R. J. Siezen, and M. Kleerebezem Functional Analysis of Three Plasmids from Lactobacillus plantarum Appl. Envir. Microbiol., March 1, 2005; 71(3): 1223 - 1230. [Abstract] [Full Text] [PDF]
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Y. G. A. EL-Osta, A. J. Hillier, and M. Dobos Construction of a combined physical and genetic map of the chromosome of Lactobacillus acidophilus ATCC 4356 and characterization of the rRNA operons Microbiology, March 1, 2005; 151(3): 875 - 892. [Abstract] [Full Text] [PDF]
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D. Dauvillee, I. S. Kinderf, Z. Li, B. Kosar-Hashemi, M. S. Samuel, L. Rampling, S. Ball, and M. K. Morell Role of the Escherichia coli glgX Gene in Glycogen Metabolism J. Bacteriol., February 15, 2005; 187(4): 1465 - 1473. [Abstract] [Full Text] [PDF]
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M. E. Schreiner and B. J. Eikmanns Pyruvate:Quinone Oxidoreductase from Corynebacterium glutamicum: Purification and Biochemical Characterization J. Bacteriol., February 1, 2005; 187(3): 862 - 871. [Abstract] [Full Text] [PDF]
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A. Benachour, C. Muller, M. Dabrowski-Coton, Y. Le Breton, J.-C. Giard, A. Rince, Y. Auffray, and A. Hartke The Enterococcus faecalis SigV Protein Is an Extracytoplasmic Function Sigma Factor Contributing to Survival following Heat, Acid, and Ethanol Treatments J. Bacteriol., February 1, 2005; 187(3): 1022 - 1035. [Abstract] [Full Text] [PDF]
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C. D. den Hengst, P. Curley, R. Larsen, G. Buist, A. Nauta, D. van Sinderen, O. P. Kuipers, and J. Kok Probing Direct Interactions between CodY and the oppD Promoter of Lactococcus lactis J. Bacteriol., January 15, 2005; 187(2): 512 - 521. [Abstract] [Full Text] [PDF]
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J. B. van Beilen, T. H. M. Smits, F. F. Roos, T. Brunner, S. B. Balada, M. Rothlisberger, and B. Witholt Identification of an Amino Acid Position That Determines the Substrate Range of Integral Membrane Alkane Hydroxylases J. Bacteriol., January 1, 2005; 187(1): 85 - 91. [Abstract] [Full Text] [PDF]
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A. Marceau, M. Zagorec, S. Chaillou, T. Mera, and M.-C. Champomier-Verges Evidence for Involvement of at Least Six Proteins in Adaptation of Lactobacillus sakei to Cold Temperatures and Addition of NaCl Appl. Envir. Microbiol., December 1, 2004; 70(12): 7260 - 7268. [Abstract] [Full Text] [PDF]
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S. H. El-Etr, S. Subbian, S. L. G. Cirillo, and J. D. Cirillo Identification of Two Mycobacterium marinum Loci That Affect Interactions with Macrophages Infect. Immun., December 1, 2004; 72(12): 6902 - 6913. [Abstract] [Full Text] [PDF]
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L. E. Hancock and M. Perego Systematic Inactivation and Phenotypic Characterization of Two-Component Signal Transduction Systems of Enterococcus faecalis V583 J. Bacteriol., December 1, 2004; 186(23): 7951 - 7958. [Abstract] [Full Text] [PDF]
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C. L. Marolda, J. Vicarioli, and M. A. Valvano Wzx proteins involved in biosynthesis of O antigen function in association with the first sugar of the O-specific lipopolysaccharide subunit Microbiology, December 1, 2004; 150(12): 4095 - 4105. [Abstract] [Full Text] [PDF]
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K. Mori, R. Bando, N. Hieda, and T. Toraya Identification of a Reactivating Factor for Adenosylcobalamin-Dependent Ethanolamine Ammonia Lyase J. Bacteriol., October 15, 2004; 186(20): 6845 - 6854. [Abstract] [Full Text] [PDF]
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