M. Kok et al., CONVERSION OF PBR322-BASED PLASMIDS INTO BROAD-HOST-RANGE VECTORS BY USING THE TN3 TRANSPOSITION MECHANISM, Journal of bacteriology, 176(21), 1994, pp. 6566-6571
We constructed a series of transposon vectors which allow efficient in
vitro gene manipulation and subsequent introduction of cloned DNA int
o a variety of gram-negative bacteria. Transfer of the cloned fragment
from these multicopy plasmids into self-transmissible broad-host-rang
e vectors is achieved in vivo, using the Tn3 transposition mechanism.
Transposition into a variety of broad-host-range plasmids proceeds eff
iciently, and the resulting recombinant plasmids can be readily transf
erred and maintained in a variety of gram-negative bacteria. The utili
ty of the transposable vectors was demonstrated by the introduction an
d expression of the lacIPOZY sequences of Escherichia coli into Pseudo
monas putida strains, allowing them to utilize lactose as a sole sourc
e of carbon and energy.