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Transformation Efficiency of EMS-induced Mutants of Streptococcus mutans of Altered Cell Shape

Department of Oral Diagnosis

T.J. MacAlister

Department of Chemical & Biological Sciences, Mount Royal College, Calgary, Alberta T2N 1N4, Canada

J.M. Tanzer

Department of Oral Diagnosis, Department of Laboratory Medicine, Schools of Dental Medicine and of Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030

Some Streptococcus mutans strains change shape from bacillary to coccal or ellipsoid form in response to the ratio of bicarbonate to potassium or of borate to potassium in growth media. So that insight into determinants of shape of these streptococci could be gained, and future genetic studies facilitated, the shapes of a series of transformable and nontransformable strains of S. mutans were studied and attempts made to isolate a mutant of augmented transformability. Several strains were mutagenized by ethylmethane sulfonate and mutants with altered colonial and cellular morphologies isolated. Cell shapes were studied by Gram stain and Nomarski interference microscopy, and by scanning and transmission electron microscopy. Diverse shape-altered mutants were isolated from seven transformable and two nontransformable strains of S. mutans. Among these, length-to-width ratios ranged from >10 to about 0.25. Regulation of timing of cell division, septum formation, or septum completion events may have been altered in these mutants. While most mutants substantially or completely lost transformability, mutant LT11 had transformation efficiency of 1.3 x 10-4 to 2.3 x 10-3, more than two to three orders of magnitude greater than its parental UA159 and the well-known transformable strain GS5(HK), respectively. There was no evidence of production of competence factor by LT11. Competence of LT11 was maintained for at least six months upon storage at -70°C, facilitating its use for genetic studies. While the morphologies of several shape-altered mutants were no longer responsive to changes of the bicarbonate/potassium, unlike those oftheir parentals, the morphology of LT11 persisted in its response to this condition. The transformation proficiency of LT11 makes it highly useful for genetic studies of S. mutans.

Journal of Dental Research, Vol. 72, No. 6, 1032-1039 (1993)
DOI: 10.1177/00220345930720060701


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