Studies on the Transfection of Phage T-1 to Corynebacterium

Song Siyang Weng Shengzhou Wu Qiaojuan ( Department of Biology )  

The transfecible of transductible phage T-l DNA to Corynebacterium has been studied. The spheroplasts were prepared in the presence of 1% gly-cine and 0.8 μg/ml penicillin G(potassium salt) and treated with 1mg/ml ly-sozyme for 10-16 hours at hypertonic media. Divalent cations such as Ca2+, Mg2+ or Mn2+ were shown to be essential for transfection. The frequency of transfection elevates remarkably in the medium containing PEG and any of the divalent cations mentioned (except Ba2+). The optimal condition for transfection was found to be 30 mM CaCl2, 16% of PEG and 1% of bovin serum albumin (BSA) at 37℃ for 5 minutes of incubation. At the optimal condition, the frequency of transfection was about 9.5×10-3p.f.u./cell and the efficiency of transfection was 2.8×106p.f.u./μg DNA.The T-l DNA could also transfect other species of Corynebacterium that usually would not be infected by this phage.

【Key Words】： Transfection Phage T-1 Corynebacterium

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