Biomaterial Database

Electrotransformation of Yersinia ruckeri by plasmid DNA. 1994

J M Cutrín, and R F Conchas, and J L Barja, and A E Toranzo

Departamento de Microbiología y Parasitología, Facultad de Biología, Universidad de Santiago de Compostela.

Yersinia ruckeri, a fish pathogenic bacterium in aquaculture, was used to evaluate the electroporation as a new transformation method for this species. DNA used for the electrotransformation were plasmids of molecular mass ranging from 2.3 kb to 33 kb, and diverse replicons. To optimize this method we used Y. ruckeri 11.29 strain (from serotype 02) and pSU2718 DNA. The best transformation efficiency (6.0 x 10(5) transformants/micrograms DNA) was obtained with 12.5 kV/cm, 25 microF, 400 omega and 2 hours of incubation after pulse. When these conditions were applied to other strains belonging to different serotypes and other plasmids, we obtained transformants in all strains assayed, but only when using low molecular weight plasmids. Plasmid vectors and resident plasmid were not modified in host strains after electrotransformation. In studies of conformation we confirmed that only circular DNA was able for transformation. The utilization of this technique for direct cloning in Y. ruckeri makes possible further studies on recombinant DNA.

UI	MeSH Term	Description	Entries
D010957	Plasmids	Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.	Episomes,Episome,Plasmid
D002455	Cell Division	The fission of a CELL. It includes CYTOKINESIS, when the CYTOPLASM of a cell is divided, and CELL NUCLEUS DIVISION.	M Phase,Cell Division Phase,Cell Divisions,Division Phase, Cell,Division, Cell,Divisions, Cell,M Phases,Phase, Cell Division,Phase, M,Phases, M
D004926	Escherichia coli	A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.	Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D013997	Time Factors	Elements of limited time intervals, contributing to particular results or situations.	Time Series,Factor, Time,Time Factor
D014169	Transformation, Bacterial	The heritable modification of the properties of a competent bacterium by naked DNA from another source. The uptake of naked DNA is a naturally occuring phenomenon in some bacteria. It is often used as a GENE TRANSFER TECHNIQUE.	Bacterial Transformation
D015007	Yersinia	A genus of gram-negative, facultatively anaerobic rod- to coccobacillus-shaped bacteria that occurs in a broad spectrum of habitats.
D015169	Colony Count, Microbial	Enumeration by direct count of viable, isolated bacterial, archaeal, or fungal CELLS or SPORES capable of growth on solid CULTURE MEDIA. The method is used routinely by environmental microbiologists for quantifying organisms in AIR; FOOD; and WATER; by clinicians for measuring patients' microbial load; and in antimicrobial drug testing.	Agar Dilution Count,Colony-Forming Units Assay, Microbial,Fungal Count,Pour Plate Count,Spore Count,Spread Plate Count,Streak Plate Count,Colony Forming Units Assay, Microbial,Colony Forming Units Assays, Microbial,Agar Dilution Counts,Colony Counts, Microbial,Count, Agar Dilution,Count, Fungal,Count, Microbial Colony,Count, Pour Plate,Count, Spore,Count, Spread Plate,Count, Streak Plate,Counts, Agar Dilution,Counts, Fungal,Counts, Microbial Colony,Counts, Pour Plate,Counts, Spore,Counts, Spread Plate,Counts, Streak Plate,Dilution Count, Agar,Dilution Counts, Agar,Fungal Counts,Microbial Colony Count,Microbial Colony Counts,Pour Plate Counts,Spore Counts,Spread Plate Counts,Streak Plate Counts
D018274	Electroporation	A technique in which electric pulses, in kilovolts per centimeter and of microsecond-to-millisecond duration, cause a loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA. Depending on the dosage, the formation of openings in the cell membranes caused by the electric pulses may or may not be reversible.	Electric Field-Mediated Cell Permeabilization,Irreversible Electroporation,Reversible Electroporation,Electropermeabilisation,Electric Field Mediated Cell Permeabilization,Electroporation, Irreversible,Electroporation, Reversible