The Experts below are selected from a list of 25437 Experts worldwide ranked by ideXlab platform
Lars M Blank - One of the best experts on this subject based on the ideXlab platform.
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida’s chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments ...
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida's chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments and flow cytometry with single-copy promoter-GFP constructs revealed a robust, constitutive behavior of these promoters, whose strengths and properties could be faithfully compared. This standardized expression device significantly extends the repertoire of tools available for reliable metabolic engineering and other genetic enhancements of P. putida.
Sebastian Zobel - One of the best experts on this subject based on the ideXlab platform.
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida’s chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments ...
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida's chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments and flow cytometry with single-copy promoter-GFP constructs revealed a robust, constitutive behavior of these promoters, whose strengths and properties could be faithfully compared. This standardized expression device significantly extends the repertoire of tools available for reliable metabolic engineering and other genetic enhancements of P. putida.
Victor De Lorenzo - One of the best experts on this subject based on the ideXlab platform.
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida’s chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments ...
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida's chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments and flow cytometry with single-copy promoter-GFP constructs revealed a robust, constitutive behavior of these promoters, whose strengths and properties could be faithfully compared. This standardized expression device significantly extends the repertoire of tools available for reliable metabolic engineering and other genetic enhancements of P. putida.
Ilaria Benedetti - One of the best experts on this subject based on the ideXlab platform.
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida’s chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments ...
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida's chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments and flow cytometry with single-copy promoter-GFP constructs revealed a robust, constitutive behavior of these promoters, whose strengths and properties could be faithfully compared. This standardized expression device significantly extends the repertoire of tools available for reliable metabolic engineering and other genetic enhancements of P. putida.
Lara Eisenbach - One of the best experts on this subject based on the ideXlab platform.
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida’s chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments ...
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tn7 based device for calibrated heterologous gene expression in pseudomonas putida
ACS Synthetic Biology, 2015Co-Authors: Sebastian Zobel, Victor De Lorenzo, Ilaria Benedetti, Lara Eisenbach, Nick Wierckx, Lars M BlankAbstract:The soil bacterium Pseudomonas putida is increasingly attracting considerable interest as a platform for advanced metabolic engineering through synthetic biology approaches. However, genomic context, gene copy number, and transcription/translation interplay often introduce considerable uncertainty to the design of reliable genetic constructs. In this work, we have established a standardized heterologous expression device in which the promoter strength is the only variable; the remaining parameters of the flow have stable default values. To this end, we tailored a mini-Tn7 delivery transposon vector that inserts the constructs in a single genomic locus of P. putida's chromosome. This was then merged with a promoter insertion site, an unvarying translational coupler, and a Downstream Location for placing the gene(s) of interest under fixed assembly rules. This arrangement was exploited to benchmark a collection of synthetic promoters with low transcriptional noise in this bacterial host. Growth experiments and flow cytometry with single-copy promoter-GFP constructs revealed a robust, constitutive behavior of these promoters, whose strengths and properties could be faithfully compared. This standardized expression device significantly extends the repertoire of tools available for reliable metabolic engineering and other genetic enhancements of P. putida.