Construction of Escherichia coli-Bifidobacterium longum shuttle vector transforming B. longum 105-A and 108-A.
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.
Matsumura
H
H
Department of Molecular Genetics, Kyoto Pharmaceutical University, Japan.
Takeuchi
A
A
Kano
Y
Y
D001644:Bifidobacterium / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D002455:Cell Division / Q000235:genetics; D015246:Deoxyribonuclease EcoRI / Q000235:genetics / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000235:genetics / Q000378:metabolism; D018274:Electroporation / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D000198:Spectinomycin / Q000494:pharmacology; D014169:Transformation, Bacterial
D000198:Spectinomycin; D009713:Nucleotidyltransferases; C022678:streptomycin 3''-adenylyltransferase; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII