A multifactorial process was used to optimize transformation of Streptococcus thermophilus by electroporation. Simple experimental designs were applied to study three, four, or five factors in eight experiments. Four qualitative factors, growth and recovering media, and plasmid and bacterial strains, were studied empirically. Eight quantitative factors, including electrical, physiological, and chemical parameters, were studied by fractional factorial designs. Effects of individual parameters as well as interactions between them were investigated and optimized. Optimization was performed for one S. thermophilus strain, ST11, and proved to work for all other tested strains of the same species. Transformation efficiencies of 9 x 10(2) to 6 x 10(5) transformants per microgram DNA were achieved, depending on the strains and vectors used.