ArticleApplied and environmental microbiology2006
Bestowing inducibility on the cloned methanol dehydrogenase promoter (PmxaF) of Methylobacterium extorquens by applying regulatory elements of Pseudomonas putida F1.
Article in Applied and environmental microbiology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed, 38 citations in OpenAlex.
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- Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.Microbial cell factories · 2025Review
- A precisely regulated Streptomyces expression system pTZYp facilitated by the strong promoter stnY and cmt operon.Biotechnology letters · 2025Article
- AFrontiers in systems biology · 2025Article
- Expression of toxic genes in Methylorubrum extorquens with a tightly repressed, cumate-inducible promoter.Antonie van Leeuwenhoek · 2023Article
- SURE editing: combining oligo-recombineering and programmable insertion/deletion of selection markers to efficiently edit the Mycoplasma pneumoniae genome.Nucleic acids research · 2022Article
- Developing methylotrophic microbial platforms for a methanol-based bioindustry.Frontiers in bioengineering and biotechnology · 2022Review
- An overview and future prospects of recombinant protein production in Bacillus subtilis.Applied microbiology and biotechnology · 2021Review
- Design and Control of Extrachromosomal Elements inACS synthetic biology · 2019Article
- Production of 2-Hydroxyisobutyric Acid from Methanol by Methylobacterium extorquens AM1 Expressing (R)-3-Hydroxybutyryl Coenzyme A-Isomerizing Enzymes.Applied and environmental microbiology · 2017Article
- Article
- New vectors for chromosomal integration enable high-level constitutive or inducible magnetosome expression of fusion proteins in Magnetospirillum gryphiswaldense.Applied and environmental microbiology · 2014Article
- Genetic and phenotypic comparison of facultative methylotrophy between Methylobacterium extorquens strains PA1 and AM1.PloS one · 2014Article
- Cumate-inducible gene expression system for sphingomonads and other Alphaproteobacteria.Applied and environmental microbiology · 2013Article
- A novel pair of inducible expression vectors for use in Methylobacterium extorquens.BMC research notes · 2013Article
- Production of functionalized polyhydroxyalkanoates by genetically modified Methylobacterium extorquens strains.Microbial cell factories · 2010Article
- Production of the chiral compound (R)-3-hydroxybutyrate by a genetically engineered methylotrophic bacterium.Applied and environmental microbiology · 2010Article
- Novel, versatile, and tightly regulated expression system for Escherichia coli strains.Applied and environmental microbiology · 2010Article
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
PmxaF is a strong methanol-inducible promoter in Methylobacterium extorquens. When this promoter is cloned in expression vectors and used to drive heterologous gene expression, methanol inducibility is either greatly reduced or entirely lost. In order to bestow inducibility upon the cloned PmxaF promoter in expression vectors, we adopted combinational methods (regulatory elements of the Pseudomonas putida F1 cym and cmt operons and Tn7 transposon system) to control reporter gene expression at the transcriptional level in M. extorquens. An operator fragment (26 nucleotides) of the cmt operon was inserted downstream of the cloned PmxaF promoter in the broad-host-range expression vector (pCHOI3). The repressor gene (cymR) located upstream of the cym operon in P. putida F1 was amplified by PCR. To avoid cellular toxicity for M. extorquens caused by the overexpression of CymR, single and/or double copies of cymR were integrated into the chromosome of M. extorquens using the mini-Tn7 transposon system. Cultures containing the chromosomally integrated cymR gene were subsequently transformed with pCHOI3 containing modified PmxaF (i.e., PmxaF plus operator). In this construct, inducibility is afforded by cumate (p-isopropylbenzoate). In this report, we describe the inducible and tightly regulated expression of heterologous genes (bgl [for beta-galactosidase], est [for esterase], and gfp [for green fluorescent protein]) in M. extorquens. This is the first documented example of an inducible/regulated heterologous gene expression system in M. extorquens.
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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.