Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster
Based on the sole information of structural genes of the 2-nitrobenzoate (2NBA) utilizing catabolic gene cluster (onbX1X2FCAR1EHJIGDBX3), 2NBA-sensing bioreporters were constructed by incorporating egfp into the onb gene cluster of Cupriavidus sp. strain ST-14. Incorporation of reporter gene in prox...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-02-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2018.00254/full |
| _version_ | 1819291557254135808 |
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| author | Satamita Deb Soumik Basu Achintya Singha Tapan K. Dutta |
| author_facet | Satamita Deb Soumik Basu Achintya Singha Tapan K. Dutta |
| author_sort | Satamita Deb |
| collection | DOAJ |
| description | Based on the sole information of structural genes of the 2-nitrobenzoate (2NBA) utilizing catabolic gene cluster (onbX1X2FCAR1EHJIGDBX3), 2NBA-sensing bioreporters were constructed by incorporating egfp into the onb gene cluster of Cupriavidus sp. strain ST-14. Incorporation of reporter gene in proximal to the hypothesized promoter region in conjunction with the disruption of the gene encoding inducer-metabolizing enzyme was turned out to be advantageous in reporter gene expression at low inducer concentration. The bioreporter strain was capable of expressing EGFP from the very 1st hour of induction and could detect 2NBA at (sub) nanomolar level exhibiting a strict specificity toward 2NBA, displaying no response to EGFP expression from its meta- and para-isomers as well as from a number of structurally related compounds. The present study is a successful demonstration of the development of a 2NBA-sensing bioreporter with respect to ease of construction, inducer specificity, and sensitivity, without prior knowledge of the associated inducer-responsive promoter-regulator elements. The present approach can be used as a model for the development of bioreporters for other environmental pollutants. |
| first_indexed | 2024-12-24T03:40:32Z |
| format | Article |
| id | doaj.art-ee1a93a8a1fa44de917e5b2a6ba5f32b |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-24T03:40:32Z |
| publishDate | 2018-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-ee1a93a8a1fa44de917e5b2a6ba5f32b2022-12-21T17:16:55ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-02-01910.3389/fmicb.2018.00254309631Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene ClusterSatamita Deb0Soumik Basu1Achintya Singha2Tapan K. Dutta3Department of Microbiology, Bose Institute, Kolkata, IndiaDepartment of Microbiology, Bose Institute, Kolkata, IndiaDepartment of Physics, Bose Institute, Kolkata, IndiaDepartment of Microbiology, Bose Institute, Kolkata, IndiaBased on the sole information of structural genes of the 2-nitrobenzoate (2NBA) utilizing catabolic gene cluster (onbX1X2FCAR1EHJIGDBX3), 2NBA-sensing bioreporters were constructed by incorporating egfp into the onb gene cluster of Cupriavidus sp. strain ST-14. Incorporation of reporter gene in proximal to the hypothesized promoter region in conjunction with the disruption of the gene encoding inducer-metabolizing enzyme was turned out to be advantageous in reporter gene expression at low inducer concentration. The bioreporter strain was capable of expressing EGFP from the very 1st hour of induction and could detect 2NBA at (sub) nanomolar level exhibiting a strict specificity toward 2NBA, displaying no response to EGFP expression from its meta- and para-isomers as well as from a number of structurally related compounds. The present study is a successful demonstration of the development of a 2NBA-sensing bioreporter with respect to ease of construction, inducer specificity, and sensitivity, without prior knowledge of the associated inducer-responsive promoter-regulator elements. The present approach can be used as a model for the development of bioreporters for other environmental pollutants.http://journal.frontiersin.org/article/10.3389/fmicb.2018.00254/fullCupriavidus sp.bioreporterinducible gene clusterenhanced green fluorescent protein2-nitrobenzoatesensitivity |
| spellingShingle | Satamita Deb Soumik Basu Achintya Singha Tapan K. Dutta Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster Frontiers in Microbiology Cupriavidus sp. bioreporter inducible gene cluster enhanced green fluorescent protein 2-nitrobenzoate sensitivity |
| title | Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster |
| title_full | Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster |
| title_fullStr | Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster |
| title_full_unstemmed | Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster |
| title_short | Development of a 2-Nitrobenzoate-Sensing Bioreporter Based on an Inducible Gene Cluster |
| title_sort | development of a 2 nitrobenzoate sensing bioreporter based on an inducible gene cluster |
| topic | Cupriavidus sp. bioreporter inducible gene cluster enhanced green fluorescent protein 2-nitrobenzoate sensitivity |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2018.00254/full |
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