A single vector system for tunable and homogeneous dual gene expression in Escherichia coli
Abstract Expression of recombinant genes can be controlled using inducible promoters. However, the most commonly used IPTG- and arabinose-inducible promoters result in an ‘all-or-nothing’ response, leading to fully induced and uninduced bacterial subpopulations. Here, we investigate whether appropri...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-024-83628-5 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841559672094457856 |
|---|---|
| author | Z. Živič L. Lipoglavšek J. Lah S. Hadži |
| author_facet | Z. Živič L. Lipoglavšek J. Lah S. Hadži |
| author_sort | Z. Živič |
| collection | DOAJ |
| description | Abstract Expression of recombinant genes can be controlled using inducible promoters. However, the most commonly used IPTG- and arabinose-inducible promoters result in an ‘all-or-nothing’ response, leading to fully induced and uninduced bacterial subpopulations. Here, we investigate whether appropriate modifications to these promoter systems can be combined into a single vector system, enabling homogenous expression of two genes of interest that can be precisely tuned using inducer concentration. We show that modifications of positive feedback loops related to inducer uptake result in homogeneous gene expression in both the T7 lactose and pBAD arabinose systems. Furthermore, these two modified systems were combined into a single vector, pRAT-sfGFP that provides the desired tunable expression of two genes of interest. Finally, we test this single-vector system as a tool for studying two-component genetic circuits, using toxin-antitoxin modules as model systems. This novel low-copy single vector expression system opens up new possibilities for investigating the function of two-component bacterial genetic circuits. |
| format | Article |
| id | doaj-art-7d0ba127484d46c1af903f95936570e7 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-7d0ba127484d46c1af903f95936570e72025-01-05T12:19:40ZengNature PortfolioScientific Reports2045-23222025-01-0115111110.1038/s41598-024-83628-5A single vector system for tunable and homogeneous dual gene expression in Escherichia coliZ. Živič0L. Lipoglavšek1J. Lah2S. Hadži3Department of Physical Chemistry, Faculty of Chemistry and Chemical Technology, University of LjubljanaChair of Microbial Diversity, Microbiomics and Biotechnology, Biotechnical Faculty, University of LjubljanaDepartment of Physical Chemistry, Faculty of Chemistry and Chemical Technology, University of LjubljanaDepartment of Physical Chemistry, Faculty of Chemistry and Chemical Technology, University of LjubljanaAbstract Expression of recombinant genes can be controlled using inducible promoters. However, the most commonly used IPTG- and arabinose-inducible promoters result in an ‘all-or-nothing’ response, leading to fully induced and uninduced bacterial subpopulations. Here, we investigate whether appropriate modifications to these promoter systems can be combined into a single vector system, enabling homogenous expression of two genes of interest that can be precisely tuned using inducer concentration. We show that modifications of positive feedback loops related to inducer uptake result in homogeneous gene expression in both the T7 lactose and pBAD arabinose systems. Furthermore, these two modified systems were combined into a single vector, pRAT-sfGFP that provides the desired tunable expression of two genes of interest. Finally, we test this single-vector system as a tool for studying two-component genetic circuits, using toxin-antitoxin modules as model systems. This novel low-copy single vector expression system opens up new possibilities for investigating the function of two-component bacterial genetic circuits.https://doi.org/10.1038/s41598-024-83628-5Dual tunable gene expressionTwo-component genetic circuitsIPTGArabinoseHigBA2Phd/Doc |
| spellingShingle | Z. Živič L. Lipoglavšek J. Lah S. Hadži A single vector system for tunable and homogeneous dual gene expression in Escherichia coli Scientific Reports Dual tunable gene expression Two-component genetic circuits IPTG Arabinose HigBA2 Phd/Doc |
| title | A single vector system for tunable and homogeneous dual gene expression in Escherichia coli |
| title_full | A single vector system for tunable and homogeneous dual gene expression in Escherichia coli |
| title_fullStr | A single vector system for tunable and homogeneous dual gene expression in Escherichia coli |
| title_full_unstemmed | A single vector system for tunable and homogeneous dual gene expression in Escherichia coli |
| title_short | A single vector system for tunable and homogeneous dual gene expression in Escherichia coli |
| title_sort | single vector system for tunable and homogeneous dual gene expression in escherichia coli |
| topic | Dual tunable gene expression Two-component genetic circuits IPTG Arabinose HigBA2 Phd/Doc |
| url | https://doi.org/10.1038/s41598-024-83628-5 |
| work_keys_str_mv | AT zzivic asinglevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT llipoglavsek asinglevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT jlah asinglevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT shadzi asinglevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT zzivic singlevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT llipoglavsek singlevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT jlah singlevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli AT shadzi singlevectorsystemfortunableandhomogeneousdualgeneexpressioninescherichiacoli |