New broad-host-range promoter probe vectors based on the plasmid RK2 replicon

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 st...

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Bibliographic Details
Published inFEMS microbiology letters Vol. 195; no. 1; pp. 91 - 96
Main Authors Santos, Pedro Miguel, Di Bartolo, Ilaria, Blatny, Janet Martha, Zennaro, Elisabetta, Valla, Svein
Format Journal Article
LanguageEnglish
Published Elsevier B.V 05.02.2001
Subjects
Broad-host-range
Plasmid
Promoter probe
RK2
Vector
Plasmid
RK2
Broad-host-range
Promoter probe
Vector
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Summary: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.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(00)00551-6