Fusion of protoplasts of Bacillus megaterium.

Fodor, Katalin; Alfoldi, L

doi:10.1073/pnas.73.6.2147

Cited by 119 publications

(57 citation statements)

References 11 publications

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“…Artificially induced fusion has been demonstrated in a number of animal cells and plant and bacterial protoplasts (Harris & Watkins, 1965;Power et al, 1970;Fodor & Alfoldi, 1976). It was reasonable to expect that yeast protoplasts could also be fused without any difficulties.…”

Section: Discussionmentioning

confidence: 99%

Fusion of Yeast Protoplasts Induced by Polyethylene Glycol

Svoboda¹

1978

Journal of General Microbiology

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Protoplasts, prepared from auxotrophic strains of Saccharomyces cerevisiae, Schizosaccharomyces pornbe and Hansenula wingei, were mixed to give intraspecific complementary combinations. Polyethylene glycol (PEG) was added to induce agglutination and fusion. Some of the fused products grew on the surface of solid minimal medium forming large vacuolated bodies. Others reverted to hybrid cells when embedded in solid minimal regeneration medium. The cytological and preliminary genetical analyses suggest a synkaryon formation and integration of genetic markers from parental strains. The frequency of intrageneric fusions assessed from the number of protoplasts growing on the surface of minimal agar was estimated to be 1 to 3 %), while the frequency of hybrid colony formation in regeneration medium was less than 1 (yo.

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Section: Discussionmentioning

confidence: 99%

Fusion of Yeast Protoplasts Induced by Polyethylene Glycol

Svoboda¹

1978

Journal of General Microbiology

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“…Their method, however, was modelled on a technique used for fusion of mammalian cells (Pontecorvo et a/., 1977) which employs a lower inolecular weight PEG (mol. wt 1540) for fusion, while the method described here is similar to the Bacillus system (Fodor & Alfoldi, 1976;Schaeffer et a/., 1976) which employs PEG 6000. With the method of Hopwood et al (1977), fused protoplasts were plated undiluted, and recombinant frequencies were determined after spore formation on confluent lawns of mycelial growth.…”

Section: Discussionmentioning

confidence: 99%

“…A lower but significant frequency of recombinants was achieved without PEG treatment. These were presumably induced to fuse by the Ca2+ ions present in medium P (Fodor & Alfoldi, 1976). Dilution of mixed protoplasts in distilled water before plating reduced the number of recombinants 100-fold.…”

Section: Genetic Recombination Induced By Protoplast Fusionmentioning

confidence: 99%

“…Polyethylene-glycol-induced protoplast (or cell) fusion has recently been demonstrated in several eukaryotic systems (Ferenczy, Kevei & Szegedi, 1975;Ann6 & Peberdy, 1976;Jones et al, 1976;Power et al, 1976;Pontecorvo, Riddle & Hales, 1977;Sipiczki & Ferenczy, 1977) and in the prokaryotic genus, Bacillus (Fodor & Alfoldi, 1976;Schaeffer, Cami & Hotchkiss, 1976). This technique appears to have potential broad utility to effect genetic recombination in prokaryotic micro-organisms, particularly in those lacking classical recombination systems.…”

Section: Introductionmentioning

confidence: 99%

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Genetic Recombination in Streptomyces fradiae by Protoplast Fusion and Cell Regeneration

Baltz

1978

Journal of General Microbiology

145

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Conditions for highly efficient genetic recombination in Streptomyces by protoplast fusion are described. Protoplasts of S. fradiae and S. griseofuscus were formed by a modification of the glycine-lysozyme-lytic enzyme method (Okanishi, Suzuki & Umezawa, 1974). Regeneration of cells from protoplasts was monitored throughout the growth cycle and was most efficient when cells of either S. fradiae or S. griseofuscus were taken from the transition phase between the exponential and stationary growth phases. Fusion of protoplasts carrying different auxotrophic or chromosomal drug-resistance markers was achieved by treatment with polyethylene glycol, and high frequencies of stable genetic recombinants were obtained. Strain development and genetic analysis of several other economically important Streptomyces species, however, has been hindered by the apparent lack of natural fertility in these strains, and the lack of transduction and transformation systems (Hopwood et al., 1973). A more general means to effect genetic recombination in Streptomyces would therefore be quite useful.Polyethylene-glycol-induced protoplast (or cell) fusion has recently been demonstrated in several eukaryotic systems (Ferenczy, Kevei & Szegedi, 1975; Ann6 & Peberdy, 1976;Jones et al., 1976;Power et al., 1976;Pontecorvo, Riddle & Hales, 1977;Sipiczki & Ferenczy, 1977) and in the prokaryotic genus, Bacillus (Fodor & Alfoldi, 1976;Schaeffer, Cami & Hotchkiss, 1976). This technique appears to have potential broad utility to effect genetic recombination in prokaryotic micro-organisms, particularly in those lacking classical recombination systems. A key factor in the utility of this technique is the capability of fused bacterial protoplasts to regenerate cell walls and thus viable cells. In this communication, I describe a highly efficient genetic recombination system for Streptomyces using protoplast fusion and efficient cell regeneration.Downloaded from www.microbiologyresearch.org by

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“…Fodor et al 16 ) reported that segregating genotypes appeared during the protoplast fusion of Bacillus megaterium. In our case, segregants did not appear after repeated transfer of the fusant from one slant to another.…”

Section: Discussionmentioning

confidence: 99%