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Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote

In prokaryotes, few studies have applied the flippase (FLP)/P1-flippase recombination target (LoxP-FRT) recombination system to switch gene expression. This study developed a new method for switching gene expression by constructing an FLP/LoxP-FRT site-specific recombination system in Escherichia co...

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Autores principales: Dan, Junhao, Deng, Huafeng, Xia, Yumei, Zhan, Yijie, Tang, Ning, Wang, Yao, Cao, Mengliang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: De Gruyter 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8919825/
https://www.ncbi.nlm.nih.gov/pubmed/35350449
http://dx.doi.org/10.1515/biol-2022-0019
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author Dan, Junhao
Deng, Huafeng
Xia, Yumei
Zhan, Yijie
Tang, Ning
Wang, Yao
Cao, Mengliang
author_facet Dan, Junhao
Deng, Huafeng
Xia, Yumei
Zhan, Yijie
Tang, Ning
Wang, Yao
Cao, Mengliang
author_sort Dan, Junhao
collection PubMed
description In prokaryotes, few studies have applied the flippase (FLP)/P1-flippase recombination target (LoxP-FRT) recombination system to switch gene expression. This study developed a new method for switching gene expression by constructing an FLP/LoxP-FRT site-specific recombination system in Escherichia coli. To this end, we placed the Nos terminator flanked by a pair of LoxP-FRT in front of enhanced green fluorescent protein (eGFP). The Nos terminator was used to block the expression of the eGFP. When a plasmid expressing FLP was available, deletion of the Nos terminator would allow expression of eGFP. The regulatory effect was demonstrated by eGFP expression. The efficiency of the gene switch was calculated as high as 89.67%. The results showed that the FLP/LoxP-FRT recombinase system could be used as a gene switch to regulate gene expression in prokaryotes. This new method for switching gene expression could simplify the gene function analysis in E. coli and other prokaryotes, as well as eukaryotes.
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spelling pubmed-89198252022-03-28 Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote Dan, Junhao Deng, Huafeng Xia, Yumei Zhan, Yijie Tang, Ning Wang, Yao Cao, Mengliang Open Life Sci Research Article In prokaryotes, few studies have applied the flippase (FLP)/P1-flippase recombination target (LoxP-FRT) recombination system to switch gene expression. This study developed a new method for switching gene expression by constructing an FLP/LoxP-FRT site-specific recombination system in Escherichia coli. To this end, we placed the Nos terminator flanked by a pair of LoxP-FRT in front of enhanced green fluorescent protein (eGFP). The Nos terminator was used to block the expression of the eGFP. When a plasmid expressing FLP was available, deletion of the Nos terminator would allow expression of eGFP. The regulatory effect was demonstrated by eGFP expression. The efficiency of the gene switch was calculated as high as 89.67%. The results showed that the FLP/LoxP-FRT recombinase system could be used as a gene switch to regulate gene expression in prokaryotes. This new method for switching gene expression could simplify the gene function analysis in E. coli and other prokaryotes, as well as eukaryotes. De Gruyter 2022-03-11 /pmc/articles/PMC8919825/ /pubmed/35350449 http://dx.doi.org/10.1515/biol-2022-0019 Text en © 2022 Junhao Dan et al., published by De Gruyter https://creativecommons.org/licenses/by/4.0/This work is licensed under the Creative Commons Attribution 4.0 International License.
spellingShingle Research Article
Dan, Junhao
Deng, Huafeng
Xia, Yumei
Zhan, Yijie
Tang, Ning
Wang, Yao
Cao, Mengliang
Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote
title Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote
title_full Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote
title_fullStr Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote
title_full_unstemmed Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote
title_short Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote
title_sort application of the flp/loxp-frt recombination system to switch the egfp expression in a model prokaryote
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8919825/
https://www.ncbi.nlm.nih.gov/pubmed/35350449
http://dx.doi.org/10.1515/biol-2022-0019
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