Abstract
To establish a DNA uptake system in coryneform bacteria, transfection of Corynebacterium glutamicum was studied. Although normal cells of C. glutamicum could not be transfected, protoplasts of C. glutamicum T-42 could be transfected with DNA of temperate phage Φcg1. Transfection was detected by the formation of plaques on a protoplast regeneration medium onto which soft agar was overlayed with transfection mixtures. Divalent cations such as CaCl2 were essential for transfection. Polyethylene glycol stimulated the efficiency of transfection about a thousand-fold. Under optimum conditions the efficiency was 9×10-2 transfected cell per input cell. Besides C. glutamicum T-42, other glutamic acid producing bacteria could also be transfected with φCG1 DNA at various frequencies.
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