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Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2
The bacterial bioluminescence system enables the generation of light by living cells without the requirement of an external luciferin. Due to the relatively low light emission, many applications of bioluminescence imaging would benefit from an increase in brightness of this system. In this report, a...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9646698/ https://www.ncbi.nlm.nih.gov/pubmed/36351939 http://dx.doi.org/10.1038/s41598-022-22068-5 |
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author | Gregor, Carola |
author_facet | Gregor, Carola |
author_sort | Gregor, Carola |
collection | PubMed |
description | The bacterial bioluminescence system enables the generation of light by living cells without the requirement of an external luciferin. Due to the relatively low light emission, many applications of bioluminescence imaging would benefit from an increase in brightness of this system. In this report, a new approach of mutagenesis and screening of the involved proteins is described that is based on the identification of mutants with improved properties under rate-limiting reaction conditions. Multiple rounds of screening in Escherichia coli resulted in the operon ilux2 that contains 26 new mutations in the fatty acid reductase complex which provides the aldehyde substrate for the bioluminescence reaction. Chromosomal integration of ilux2 yielded an autonomously bioluminescent E. coli strain with sixfold increased brightness compared to the previously described ilux operon. The ilux2 strain produces sufficient signal for the robust detection of individual cells and enables highly sensitive long-term imaging of bacterial propagation without a selection marker. |
format | Online Article Text |
id | pubmed-9646698 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-96466982022-11-15 Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 Gregor, Carola Sci Rep Article The bacterial bioluminescence system enables the generation of light by living cells without the requirement of an external luciferin. Due to the relatively low light emission, many applications of bioluminescence imaging would benefit from an increase in brightness of this system. In this report, a new approach of mutagenesis and screening of the involved proteins is described that is based on the identification of mutants with improved properties under rate-limiting reaction conditions. Multiple rounds of screening in Escherichia coli resulted in the operon ilux2 that contains 26 new mutations in the fatty acid reductase complex which provides the aldehyde substrate for the bioluminescence reaction. Chromosomal integration of ilux2 yielded an autonomously bioluminescent E. coli strain with sixfold increased brightness compared to the previously described ilux operon. The ilux2 strain produces sufficient signal for the robust detection of individual cells and enables highly sensitive long-term imaging of bacterial propagation without a selection marker. Nature Publishing Group UK 2022-11-09 /pmc/articles/PMC9646698/ /pubmed/36351939 http://dx.doi.org/10.1038/s41598-022-22068-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Gregor, Carola Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
title | Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
title_full | Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
title_fullStr | Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
title_full_unstemmed | Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
title_short | Generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
title_sort | generation of bright autobioluminescent bacteria by chromosomal integration of the improved lux operon ilux2 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9646698/ https://www.ncbi.nlm.nih.gov/pubmed/36351939 http://dx.doi.org/10.1038/s41598-022-22068-5 |
work_keys_str_mv | AT gregorcarola generationofbrightautobioluminescentbacteriabychromosomalintegrationoftheimprovedluxoperonilux2 |