CONVERSION OF PBR322-BASED PLASMIDS INTO BROAD-HOST-RANGE VECTORS BY USING THE TN3 TRANSPOSITION MECHANISM

Citation
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
Citations number
25
Categorie Soggetti
Microbiology
Journal title
ISSN journal
00219193
Volume
176
Issue
21
Year of publication
1994
Pages
6566 - 6571
Database
ISI
SICI code
0021-9193(1994)176:21<6566:COPPIB>2.0.ZU;2-3
Abstract
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.