Novel plasmid vectors for gene cloning in Pseudomonas.
Novel host-vector systems have been developed for gene cloning in the metabolically versatile bacterial genus Pseudomonas. We found that a new Pseudomonas strain, Pseudomonas flavida IF-4, isolated from soil, carried two small cryptic plasmids, named pNI10 and pNI20. They were multi-copy, but not self-transmissible, and the genome size was 3.7 kb for pNI10 and 2.9 kb for pNI20. Several types of cloning vectors containing a kanamycin or streptomycin resistance (Kmr or Smr) gene were constructed from pNI10 and pNI20. These plasmid vectors were efficiently transformed into several strains of Pseudomonas at a frequency up to 4 x 10(5) transformants per 1 microgram plasmid DNA by the usual competent cell method. The vectors derived from pNI10 replicated not only in Pseudomonas but also in some other Gram-negative enteric bacteria such as Escherichia coli, Enterobacter aerogenes, and Proteus mirabilis.
Itoh
N
N
Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Fukui University.
Koide
Y
Y
Fukuzawa
H
H
Hirose
S
S
Inukai
T
T
D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods*; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors; D008854:Microscopy, Electron; D010957:Plasmids; D011549:Pseudomonas / Q000145:classification / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D016958:Pseudomonas putida / Q000235:genetics; D015183:Restriction Mapping
D003470:Culture Media; D004269:DNA, Bacterial