title: Transformation of alternan-producing strains of Leuconostoc by electroporation.
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 Omega and 10 kV cm(-1), resulting in transformation efficiencies of up to 3.5 x 10(4) per microg 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.
authors.affiliation: Fermentation Biotechnology Research Unit, National Center for Agricultural Utilization Research, USDA, Agricultural Research Service, Peoria, IL 61604, USA. leathetd@ncaur.usda.gov
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Leathers
authors.forename: James D
authors.initials: JD
authors.lastname: Jones
authors.forename: Herb A
authors.initials: HA
authors.lastname: Wyckoff
mesh_terms: D001709:Biotechnology / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D018274:Electroporation / Q000379:methods*; D004996:Ethidium / Q000494:pharmacology; D005822:Genetic Vectors; D005936:Glucans / Q000096:biosynthesis*; D007934:Leuconostoc / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial
chemical_list: D005936:Glucans; C439358:alternan; D004247:DNA; D004996:Ethidium