Open AccessNew broad‐host‐range promoter probe vectors based on the plasmid RK2 replicon
Author(s) -
Santos Pedro Miguel,
Bartolo Ilaria,
Blatny Janet Martha,
Zennaro Elisabetta,
Valla Svein
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10503.x
Subject(s) - biology , plasmid , replicon , shuttle vector , genetics , reporter gene , pseudomonas putida , escherichia coli , lac operon , pseudomonas fluorescens , gene , expression vector , inducer , microbiology and biotechnology , vector (molecular biology) , bacteria , gene expression , recombinant dna
Broad‐host‐range plasmid RK2‐based promoter probe vectors with a known nucleotide sequence were constructed. In the absence of an upstream promoter, the expression of two tested reporter genes ( luc and lacZ ) in Escherichia coli was virtually zero, while insertion of the Ptrc promoter resulted in strong inducer‐dependent expression. The lacZ‐ based vectors were mobilized into Pseudomonas fluorescens ST, Pseudomonas putida KT2442, Sphingomonas spp. and Burkholderia spp. LB400, and expression analyses indicated that the properties observed in E. coli are maintained across the species barriers. In addition, the previously established knowledge of RK2 molecular biology allows easy manipulations of features such as plasmid copy number, further extending the application potential of the vectors.