Bioscience Biotechnology and Biochemistry · 1997 · 84 citations · 0 references
Life SciencesBiotechnologyMicrobiologyBacteriologyGenetic EngineeringMicrobial BiotechnologyMicrobial Cell EngineeringPlasmid BiologyPlasmid GeneticsStrain ImprovementTransgenic Organism ProductionTransgenic OrganismsBacterial Genome EngineeringBacterial Plasmid ConstructionBacterial TransformationEscherichia ColiGenetic Engineering Of MicrobesShuttle Vector ConstructionShuttle VectorsTransgenic Microbial Strains
A shuttle vector, pBLES100, was constructed by cloning a Bifidobacterium longum plasmid and a gene encoding spectinomycin adenyltransferase AAD(9) from Enterococcus faecalis into the Escherichia coli vector pBR322. Stable transformants with this plasmid were obtained with an efficiency of 2.2 x 10(4) transformants/microgram DNA or 6.9 x 10(-5) transformants/cell/microgram DNA under the optimal conditions of 10.0 kV/cm, 200 omega, and 25 microF, using B. longum 105-A harvested at late log phase of growth.