Agricultural and Biological Chemistry
Online ISSN : 1881-1280
Print ISSN : 0002-1369
ISSN-L : 0002-1369
Interspecific Electrofusion between Protoplasts of Streptomyces antibioticus and Streptomyces fradiae
Tokumitsu OKAMURAShinji NAGATAHamo MISONOSusumu NAGASAKI
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1988 Volume 52 Issue 6 Pages 1433-1438

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Abstract

The interspecific electrofusion between protoplasts of Streptomyces antibioticus a20 (Ala-) and S. fradiae f2 (His-) was examined. The pulse intensities required for the breakdown of half of the protoplasts were 7kV/cm for S. antibioticus a20 and 5kV/cm for S. fradiae f2. The optimum pulse intensity for the fusion was found to be 6 kV/cm. When the density of a protoplast suspension was 1 × 108 protoplasts/ml in a 0.3M sucrose solution containing 2.5mM MgCl2 and CaCl2, the maximum fusion frequency of 2.7% was obtained on the application of one square field pulse of 6kV/cm for 20μsec after dielectrophoresis in an alternating field of 800V/cm (1 MHz) for 60 sec. The fusion frequency was much higher than that obtained on polyethyleneglycol-induced fusion (0.15%). This is the first case of electrofusion of Streptomyces protoplasts, which are as small as those of some bacteria.

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