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Abstract
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.
Highlights
In prokaryotes, few studies have applied the flippase (FLP)/P1-flippase recombination target (LoxPFRT) recombination system to switch gene expression
The FLP/LoxP-FRT recombination system was used as a gene switch, controlling the temporal and spatial specificity to precisely regulate target gene expression
The FLP recombination acted as the “gene key” while Nos terminator was the “gene lock” that hindered the expression of enhanced green fluorescent protein (eGFP)
Summary
Abstract: In prokaryotes, few studies have applied the flippase (FLP)/P1-flippase recombination target (LoxPFRT) 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 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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