ArticleGene2007
Functional expression of the Flp recombinase in Mycobacterium bovis BCG.
Article in Gene, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 30 citations in OpenAlex.
- Modular toolkit to facilitate molecular manipulations in mycobacteria.Journal of bacteriology · 2026Article
- Uncovering Insights Into the Biology of Mycobacterium tuberculosis Using Genetic Tools.MicrobiologyOpen · 2025Review
- Evolutionary Trajectories of Methionine Metabolism in Mycobacterium and Its Application to Engineer a Vitamin B12 Whole-Cell Ribosensor.Microbial biotechnology · 2025Article
- Genetic Lineage Tracing of Non-cardiomyocytes in Mice.Methods in molecular biology (Clifton, N.J.) · 2021Article
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- CRISPR-Cas12a-Assisted Recombineering in Bacteria.Applied and environmental microbiology · 2017Article
- Mob/oriT, a mobilizable site-specific recombination system for unmarked genetic manipulation in Bacillus thuringiensis and Bacillus cereus.Microbial cell factories · 2016Article
- Efficient and simple generation of multiple unmarked gene deletions in Mycobacterium smegmatis.Scientific reports · 2016Article
- Rhomboids of Mycobacteria: characterization using an aarA mutant of Providencia stuartii and gene deletion in Mycobacterium smegmatis.PloS one · 2012Article
- Article
- Xer site-specific recombination, an efficient tool to introduce unmarked deletions into mycobacteria.Applied and environmental microbiology · 2010Article
- Application of the Saccharomyces cerevisiae FLP/FRT recombination system in filamentous fungi for marker recycling and construction of knockout strains devoid of heterologous genes.Applied and environmental microbiology · 2010Article
- Rapid evaluation of the mycobactericidal efficacy of disinfectants in the quantitative carrier test EN 14563 by using fluorescent Mycobacterium terrae.Applied and environmental microbiology · 2010Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Mycobacteria contain a large number of redundant genes whose functions are difficult to analyze in mutants because there are only two efficient antibiotic resistance genes available for allelic exchange experiments. Sequence-specific recombinbases such as the Flp recombinase can be used to excise resistance markers. Expression of the flp(e) gene from Saccharomyces cerevisiae is functional for this purpose in fast-growing Mycobacterium smegmatis but not in slow-growing mycobacteria such as M. bovis BCG or M. tuberculosis. We synthesized the flp(m) gene by adapting the codon usage to that preferred by M. tuberculosis. This increased the G+C content from 38% to 61%. Using the synthetic flp(m) gene, the frequency of removal of FRT-hyg-FRT cassette from the chromosome by the Flp recombinase was increased by more than 100-fold in M. smegmatis. In addition, 40% of all clones of M. bovis BCG had lost the hyg resistance cassette after transient expression of the flp(m) gene. Sequencing of the chromosomal DNA showed that excision of the FRT-hyg-FRT cassette by Flp was specific. These results show that the flp(m) encoded Flp recombinase is not only an improved genetic tool for M. smegmatis, but can also be used in slow growing mycobacteria such as M. tuberculosis for constructing unmarked mutations. Other more sophisticated applications in mycobacterial genetics would also profit from the improved Flp/FRT system.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.