This study describes a broad host transformation protocol that enables the uptake of plasmid DNA into 10 different species of Bifidobacterium, some of which have never been transformed before. The vector pNC7 (4·9 kb) was used to optimize the electroporation protocol. Transformation efficiencies ranged from 3·6×10−1 to 1·2×105 transformations per μg DNA. The impact of growth medium composition and electric field strength on transformation efficiency were independently optimized. Electrocompetent cells were grown in Iwata medium broth enriched with ActilightRP 16%, harvested during the early exponential growth phase, and pulsed at 12·5 kV cm−1, 100 Ω and 25 μF.
An efficient transformation system for Bifidobacterium spp / Rossi, Maddalena; P., Brigidi; D., Matteuzzi. - In: LETTERS IN APPLIED MICROBIOLOGY. - ISSN 0266-8254. - ELETTRONICO. - 24:(1996), pp. 33-36.
An efficient transformation system for Bifidobacterium spp.
ROSSI, Maddalena;
1996
Abstract
This study describes a broad host transformation protocol that enables the uptake of plasmid DNA into 10 different species of Bifidobacterium, some of which have never been transformed before. The vector pNC7 (4·9 kb) was used to optimize the electroporation protocol. Transformation efficiencies ranged from 3·6×10−1 to 1·2×105 transformations per μg DNA. The impact of growth medium composition and electric field strength on transformation efficiency were independently optimized. Electrocompetent cells were grown in Iwata medium broth enriched with ActilightRP 16%, harvested during the early exponential growth phase, and pulsed at 12·5 kV cm−1, 100 Ω and 25 μF.
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