Transformation of Streptococcus bovis protoplasts by plasmid DNA.
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.
Mareková
M
M
Institute of Animal Physiology, Slovak Academy of Science, Kosice, Slovakia.
Kmet
V
V
Javorský
P
P
D000818:Animals; D002417:Cattle; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005821:Genetic Techniques; D010957:Plasmids / Q000235:genetics*; D011092:Polyethylene Glycols; D011523:Protoplasts; D012417:Rumen / Q000382:microbiology; D016985:Streptococcus bovis / Q000235:genetics*; D013395:Sucrose; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D011092:Polyethylene Glycols; D013395:Sucrose