Fusion of Erwinia chrysanthemi spheroplasts in an electric fields

A new approach has been elaborated for electrofusion of Erwinia chrysanthemi spheroplasts. The new approach consists of superimposition of high voltage impulses on the pellet of tightly contacting cells in the course of centrifugation. The mixture of spheroplasts of two genetically marked strains wa...

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Bibliographic Details
Published inMolekulârnaâ genetika, mikrobiologiâ i virusologiâ no. 5; p. 30
Main Authors Iakovenko, K N, Khendogiĭ, N V, Sukharev, S I, Abidor, I G, Troitskiĭ, N A
Format Journal Article
LanguageRussian
Published Russia (Federation) 01.05.1990
Subjects
Bacteriological Techniques
Culture Media
Electricity
Erwinia - genetics
Erwinia - physiology
Erwinia - radiation effects
Polyethylene Glycols - pharmacology
Recombination, Genetic
Spheroplasts - physiology
Spheroplasts - radiation effects
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Summary:A new approach has been elaborated for electrofusion of Erwinia chrysanthemi spheroplasts. The new approach consists of superimposition of high voltage impulses on the pellet of tightly contacting cells in the course of centrifugation. The mixture of spheroplasts of two genetically marked strains was placed into the special centrifuge chambers and spinned for 15 min at 2500 g to get a compressed pellet between chamber electrodes. Three successive pulses of 6.6 kv/cm amplitude and 30 microseconds duration were applied to spheroplast pellet during centrifugation. Fusion products were viable and after plating on the surface of hypertonic medium regenerated to the rod forms. As a result, the hybrid clones carrying the markers of both parents were isolated.
ISSN:0208-0613