scholarly journals PBAD-Based Shuttle Vectors for Functional Analysis of Toxic and Highly Regulated Genes in Pseudomonas and Burkholderia spp. and Other Bacteria

2008 ◽  
Vol 74 (23) ◽  
pp. 7422-7426 ◽  
Author(s):  
Dongru Qiu ◽  
F. Heath Damron ◽  
Takehiko Mima ◽  
Herbert P. Schweizer ◽  
Hongwei D. Yu
Keyword(s):  
Functional Analysis ◽  
Host Range ◽  
Expression Vectors ◽  
Broad Host Range ◽  
Shuttle Vectors ◽  
Conditional Expression

ABSTRACT We report the construction of a series of Escherichia-Pseudomonas broad-host-range expression vectors utilizing the PBAD promoter and the araC regulator for routine cloning, conditional expression, and analysis of tightly controlled and/or toxic genes in pseudomonads.

pIJ101, a multi-copy broad host-range Streptomyces plasmid: Functional analysis and development of DNA cloning vectors

1982 ◽  
Vol 185 (2) ◽  
pp. 223-238 ◽  
Author(s):  
Tobias Kieser ◽  
David A. Hopwood ◽  
Helen M. Wright ◽  
Charles J. Thompson
Keyword(s):  
Functional Analysis ◽  
Host Range ◽  
Broad Host Range ◽  
Cloning Vectors ◽  
Dna Cloning

Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors

Gene ◽  
1983 ◽  
Vol 26 (2-3) ◽  
pp. 273-282 ◽  
Author(s):  
Miroslawa M. Bagdasarian ◽  
Egon Amann ◽  
Rudolf Lurz ◽  
Beate Rückert ◽  
Michael Bagdasarian
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Putida ◽  
Host Range ◽  
Expression Vectors ◽  
Broad Host Range ◽  
Tac Promoter

scholarly journals Broad-Host-Range Expression Vectors with Tightly Regulated Promoters and Their Use To Examine the Influence of TraR and TraM Expression on Ti Plasmid Quorum Sensing

2008 ◽  
Vol 74 (16) ◽  
pp. 5053-5062 ◽  
Author(s):  
Sharik R. Khan ◽  
Jennifer Gaines ◽  
R. Martin Roop ◽  
Stephen K. Farrand
Keyword(s):  
Host Range ◽  
Gene Activation ◽  
Ti Plasmid ◽  
Expression Vectors ◽  
Multiple Cloning Site ◽  
Broad Host Range ◽  
Precise Control ◽  
Broad Host Range Plasmid ◽  
Cloned Gene ◽  
Ndei Site

ABSTRACT Experiments requiring strong repression and precise control of cloned genes can be difficult to conduct because of the relatively high basal level of expression of currently employed promoters. We report the construction of a family of vectors that contain a reengineered lacI q-lac promoter-operator complex in which cloned genes are strongly repressed in the absence of inducer. The vectors, all based on the broad-host-range plasmid pBBR1, are mobilizable and stably replicate at moderate copy number in representatives of the alpha- and gammaproteobacteria. Each vector contains a versatile multiple cloning site that includes an NdeI site allowing fusion of the cloned gene to the initiation codon of lacZα. In each tested bacterium, a uidA reporter fused to the promoter was not expressed at a detectable level in the absence of induction but was inducible by 10- to 100-fold, depending on the bacterium. The degree of induction was controllable by varying the concentration of inducer. When the vector was tested in Agrobacterium tumefaciens, a cloned copy of the traR gene, the product of which is needed at only a few copies per cell, did not confer activity under noninducing conditions. We used this attribute of very tight and variably regulatable control to assess the relative amounts of TraR required to activate the Ti plasmid conjugative transfer system. We identified levels of induction that gave wild-type transfer frequencies, as well as levels that induced correspondingly lower frequencies of transfer. We also used this system to show that the antiactivator TraM sets the level of intracellular TraR required for tra gene activation.

Novel broad host range shuttle vectors for expression in Escherichia coli, Bacillus subtilis and Pseudomonas putida

2012 ◽  
Vol 161 (2) ◽  
pp. 71-79 ◽  
Author(s):  
Sonja Christina Troeschel ◽  
Stephan Thies ◽  
Olga Link ◽  
Catherine Isabell Real ◽  
Katja Knops ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Pseudomonas Putida ◽  
Host Range ◽  
Broad Host Range ◽  
Shuttle Vectors ◽  
Expression In Escherichia Coli

scholarly journals Plasmid Vectors for Xylella fastidiosa Utilizing a Toxin-Antitoxin System for Stability in the Absence of Antibiotic Selection

2016 ◽  
Vol 106 (8) ◽  
pp. 928-936 ◽  
Author(s):  
Lindsey P. Burbank ◽  
Drake C. Stenger
Keyword(s):  
Copy Number ◽  
Xylella Fastidiosa ◽  
Expression Vectors ◽  
Broad Host Range ◽  
In Planta ◽  
Plasmid Vectors ◽  
Antibiotic Selection ◽  
Shuttle Vectors ◽  
Range Vector

The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacterial genetics but there are only a limited number of plasmid vectors available that replicate in X. fastidiosa, and even fewer that are retained without antibiotic selection. Two plasmids are described here that show stable replication in X. fastidiosa and are effective for gene complementation both in vitro and in planta. Plasmid maintenance is facilitated by incorporation of the PemI/PemK plasmid addiction system, consisting of PemK, an endoribonuclease toxin, and its cognate antitoxin, PemI. Vector pXf20pemIK utilizes a native X. fastidiosa replication origin as well as a high-copy-number pUC origin for propagation in Escherichia coli cloning strains. Broad-host-range vector pBBR5pemIK is a medium- to low-copy-number plasmid based on the pBBR1 backbone. Both plasmids are maintained for extended periods of time in the absence of antibiotic selection, as well as up to 14 weeks in grapevine, without affecting bacterial fitness. These plasmids present an alternative to traditional complementation and expression vectors which rely on antibiotic selection for plasmid retention.

Broad-host range expression vectors containing manipulatedmeta-cleavage pathway regulatory elements of the TOL plasmid

FEBS Letters ◽  
1988 ◽  
Vol 226 (2) ◽  
pp. 241-246 ◽  
Author(s):  
Juan L. Ramos ◽  
Manuel Gonzalez-Carrero ◽  
Kenneth N. Timmis
Keyword(s):  
Host Range ◽  
Regulatory Elements ◽  
Expression Vectors ◽  
Broad Host Range ◽  
Tol Plasmid
Sign in / Sign up

Export Citation Format

Share Document