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Transformation of Streptococcus bovis protoplasts by plasmid DNA
M Mareková1, V Kmet, P Javorský
1Institute of Animal Physiology, Slovak Academy of Science, Kosice, Slovakia.
Letters in Applied Microbiology
|February 1, 1996
Abstract:
The transformation and subsequent regeneration of ruminal strain Streptococcus bovis AO24/85 protoplasts by plasmid DNA was studied. The best stabilizer for regeneration of protoplasted cells was 5% sucrose in the regeneration medium and in the agar plates. Optimal concentration of polyethylene glycol 6000 in the transformation medium was 25% for both plasmids tested. Addition of Ca2+ and Mg2+ ions (2.5 mmol l-1) to the transformation medium increased the proportion of regenerated cells. Transformation frequencies were 3 x 10(3) transformants per microgram of pNZ12 and 2.4 x 10(2) per microgram of pJK108, respectively.