Transformation of bacteria by electroporation

Chassy, Bruce M.; Mercenier, Annick; Flickinger, J L

doi:10.1016/0167-7799(88)90025-x

Cited by 129 publications

(58 citation statements)

References 29 publications

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“…Electroporation is used as an efficient transformation technique for gram-positive and gram-negative bacteria (Chassy et al 1988;Dower et al 1988;Fiedler and Wirth 1988;Tryfona and Bustard 2005). Mechanisms of gene electrotransfer were studied extensively, among which surface binding and diffusion through electropores, effective electric pulse parameters, and the effect of DNA topology on transformation efficiency were investigated (Xie et al 1990;.…”

Section: Electroporation Of Bacteria and Yeastmentioning

confidence: 99%

Electroporation in Biological Cell and Tissue: An Overview

Kandušer

Miklavčič²

2008

Electrotechnologies for Extraction From Food Plants and Biomaterials

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In this chapter, basics and mechanisms of electroporation are presented. Most important electric pulse parameters for electroporation efficiency for different applications that involve introduction of small molecules and macromolecules into the cell or cell membrane electrofusion are described. In all these applications, cell viability has to be preserved. However, in some biotechnological applications, such as liquid food sterilization or water treatment, electroporation is used as a method for efficient cell killing. For all the applications mentioned above, besides electric pulse parameters, other factors, such as electroporation medium composition and osmotic pressure, play significant roles in electroporation effectiveness. For controlled use of the method in all applications, the basic mechanisms of electroporation need to be known. The phenomenon was studied from the single-cell level and dense cell suspension that represents a simplified homogenous tissue model, to complex biological tissues. In the latter, different cell types and electric conductivity that change during the course of electric pulse application can significantly affect the effectiveness of the treatment. For such a complex situation, the design and use of suitable electrodes and theoretical modeling of electric field distribution within the tissue are essential. Electroporation as a universal method applicable to different cell types is used for different purposes. In medicine it is used for electrochemotherapy and genetherapy. In biotechnology it is used for water and liquid food sterilization and for transfection of bacteria, yeast, plant protoplast, and intact plant tissue. Understanding the phenomenon of electroporation, its mechanisms and optimization of all the parameters that affect electroporation is a prerequisite for successful treatment. In addition to the parameters mentioned above, different biological characteristics of treated cell affect the outcome of the treatment. Electroporation, gene electrotransfer and electrofusion are affected by cell membrane fluidity, cytoskeleton, and the presence of the cell wall in bacteria yeast and plant cells. Thus, electroporation parameters need to be specifically optimized for different cell types.

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Section: Electroporation Of Bacteria and Yeastmentioning

confidence: 99%

Electroporation in Biological Cell and Tissue: An Overview

Kandušer

Miklavčič²

2008

Electrotechnologies for Extraction From Food Plants and Biomaterials

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“…The ability to electroporate protoplasts, spheroplasts and intact cells has advanced microbiological studies in organisms where other transformation procedures have (Chassy et al, 1988). However, the generation of cells lacking cell walls can be labour-intensive, timeconsuming and difficult to reproduce.…”

Section: Introductionmentioning

confidence: 99%

Development of a P1 phagemid system for the delivery of DNA into Gram-negative bacteria

et al. 2002

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The inability to transform many clinically important Gram-negative bacteria has hampered genetic studies addressing the mechanism of bacterial pathogenesis. This report describes the development and construction of a delivery system utilizing the broad-host-range transducing bacteriophage P1. The phagemids used in this system contain a P1 pac initiation site to package the vector, a P1 lytic replicon to generate concatemeric DNA, a broad-hostrange origin of replication and an antibiotic-resistance determinant to select bacterial clones containing the recircularized phagemid. Phagemid DNA was successfully introduced by infection and stably maintained in members of the families Enterobacteriaceae (Escherichia coli, Shigella flexneri, Shigella dysenteriae, Klebsiella pneumoniae and Citrobacter freundii ) and Pseudomonadaceae (Pseudomonas aeruginosa). In addition to laboratory strains, these virions were used successfully to deliver phagemids to a number of strains isolated from patients. This ability to deliver genetic information to wild-type strains raises the potential for use in antimicrobial therapies and DNA vaccine development.

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“…Routinely, E. coli strains were transformed by the calcium chloride-rubidium chloride method, as described previously (27). For transformation with complex ligation mixes (three-and four-way ligations), cells were transformed by electroporation (14) (electrotransformation [9]). Electrotransformation was also used for E. coli E274, SA500, SA599, and SA1287, which are only poorly transformable by other procedures.…”

mentioning

confidence: 99%

Carbohydrate utilization in Streptococcus thermophilus: characterization of the genes for aldose 1-epimerase (mutarotase) and UDPglucose 4-epimerase

Poolman¹,

Royer²,

Mainzer³

et al. 1990

J Bacteriol

109

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The complete nucleotide sequences of the genes encoding aldose 1-epimerase (mutarotase) (galM) and UDPglucose 4-epimerase (galE) and flanking regions of Streptococcus thermophilus have been determined. Both genes are located immediately upstream of the S. thermophilus lac operon. To facilitate the isolation of galE, a special polymerase chain reaction-based technique was used to amplify the region upstream of galM prior to cloning. The galM protein was homologous to the mutarotase of Acinetobacter calcoaceticus, whereas the galE protein was homologous to UDPglucose 4-epimerase of Escherichia coli and Streptomyces lividans. The amino acid sequences of galM and galE proteins also showed significant similarity with the carboxy-terminal and amino-terminal domains, respectively, of UDPglucose 4-epimerase from Kluyveromyces lactis and Saccharomyces cerevisiae, suggesting that the yeast enzymes contain an additional, yet unidentified (mutarotase) activity. In accordance with the open reading frames of the structural genes, galM and galE were expressed as polypeptides with apparent molecular masses of 39 and 37 kilodaltons, respectively. Significant activities of mutarotase and UDPglucose 4-epimerase were detected in lysates of E. coli cells containing plasmids encoding galM and galE. Expression of galE in E. coli was increased 300-fold when the gene was placed downstream of the tac promoter. The gene order for the gal-lac gene cluster of S. thermophilus is galE-gahM-lacS-lacZ. The flanking regions of these genes were searched for consensus promoter sequences and further characterized by primer extension analysis. Analysis of mRNA levels for the gal and lac genes in S. thermophilus showed a strong reduction upon growth in medium containing glucose instead of lactose. The activities of the lac (lactose transport and Il-galactosidase) and gal (UDPglucose 4-epimerase) proteins of lactose-and glucose-grown S. thermophilus cells matched the mRNA levels.Streptococcus thermophilus transports lactose by means of a proton motive force-linked mechanism (33). Lactose enters the cell as a free sugar, and the disaccharide is hydrolyzed into glucose and galactose by P-galactosidase (20,33). Glucose enters the glycolytic pathway, whereas in the presence of excess lactose, the galactose moiety of lactose is excreted into the medium (39).The lac genes of S. thermophilus have recently been cloned, sequenced, and partially characterized (20, 33; C. J. Schroeder, C. Robert, G. Lenzen, L. L. McKay, and A.Mercenier, submitted for publication). The lactose transport gene (lacS) encodes a 69,454-dalton (Da) protein consisting of an amino-terminal domain with homology to the melibiose carrier of Escherichia coli and a carboxy-terminal domain with homology to enzyme III or enzyme III domains of various phosphoenolpyruvate-dependent phosphotransferase systemns from gram-positive and gram-negative organisms. A similar transport protein has been found in Lactobacillus bulgaricus (33,38), and the function(s) of the different domains of the transport...

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Transformation of bacteria by electroporation

Cited by 129 publications

References 29 publications

Electroporation in Biological Cell and Tissue: An Overview

Electroporation in Biological Cell and Tissue: An Overview

Development of a P1 phagemid system for the delivery of DNA into Gram-negative bacteria

Carbohydrate utilization in Streptococcus thermophilus: characterization of the genes for aldose 1-epimerase (mutarotase) and UDPglucose 4-epimerase

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