Abstract
Alternan-producing Leuconostoc mesenteroides strain NRRL B-1355 and its glucansucrase-negative derivative NRRL B-21414 were transformed by electroporation using four Gram positive-Gram negative shuttle vectors. Optimal conditions were 400 Ω and 10 kV cm−1, resulting in transformation efficiencies of up to 3.5×104 per μg DNA. Relatively low copy numbers and native plasmids made it difficult to visualize the introduced plasmids on ethidium bromide-stained gels and, in some cases, on blot hybridizations. However, PCR analysis indicated that 95% of putative transformants carried plasmid sequences. Direct colony PCR was shown to work well for this system and also for transformants of L. mesenteroides subsp. cremoris.
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Leathers, T.D., Jones, J.D. & Wyckoff, H.A. Transformation of alternan-producing strains of Leuconostoc by electroporation. Biotechnology Letters 26, 1119–1124 (2004). https://doi.org/10.1023/B:BILE.0000035482.41491.7e
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DOI: https://doi.org/10.1023/B:BILE.0000035482.41491.7e