PREPARATION OF SPHEROPLASTS FROM VIBRIO COMMA

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Abstract
Chatterjee , B. R. (Baylor University College of Medicine, Houston, Texas), and Robert P. Williams . Preparation of spheroplasts from Vibrio comma . J. Bacteriol. 85: 838–841. 1963.—Spheroplasts were prepared from several strains of Vibrio comma by lysozyme treatment combined with freezing and thawing of the organisms. The optimal concentration of lysozyme was 50 μg/ml, although some spheroplasts formed at a concentration of 10 μg/ml. Higher concentrations (200 μg/ml) caused lysis of cells along with spheroplast formation. Treatment was carried out in broth cultures containing 15% sucrose, and if the osmotic tension was lowered the spheroplasts lysed. Some motile, spherical cells were present in every preparation. Addition of 3% glycine to broth cultures resulted in rapid transformation of the vibrios into large, spherical bodies. However, these were actively motile, and were not sensitive to a lower osmotic tension. Therefore, they could not be considered as spheroplasts.