ArticleApplied and environmental microbiology2006
Multicopy integration and expression of heterologous genes in Methylobacterium extorquens ATCC 55366.
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 17 papers.
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17 citing papers in PubMed, 34 citations in OpenAlex.
- Engineering a Fusion Protein Biomaterial Based on SpyTag/SpyCatcher Bioconjugation of Elastin and Collagen Synthetic Proteins.ACS omega · 2025Article
- Heterologous production of 3-hydroxypropionic acid in Methylorubrum extorquens by introducing the mcr gene via a multi-round chromosomal integration system based on cre-lox71/lox66 and transposon.Microbial cell factories · 2024Article
- Biosynthesis of polyhydroxybutyrate byFrontiers in plant science · 2024Article
- Levulinic Acid-Inducible and Tunable Gene Expression System forFrontiers in bioengineering and biotechnology · 2021Article
- GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep.Transboundary and emerging diseases · 2019Article
- Article
- Production of 2-Hydroxyisobutyric Acid from Methanol by Methylobacterium extorquens AM1 Expressing (R)-3-Hydroxybutyryl Coenzyme A-Isomerizing Enzymes.Applied and environmental microbiology · 2017Article
- Metabolic footprint of epiphytic bacteria on Arabidopsis thaliana leaves.The ISME journal · 2016Article
- Parallel and Divergent Evolutionary Solutions for the Optimization of an Engineered Central Metabolism in Methylobacterium extorquens AM1.Microorganisms · 2015Article
- New vectors for chromosomal integration enable high-level constitutive or inducible magnetosome expression of fusion proteins in Magnetospirillum gryphiswaldense.Applied and environmental microbiology · 2014Article
- Tuning the dials of Synthetic Biology.Microbiology (Reading, England) · 2013Review
- Application of the bacteriophage Mu-driven system for the integration/amplification of target genes in the chromosomes of engineered Gram-negative bacteria--mini review.Applied microbiology and biotechnology · 2011Review
- Production of functionalized polyhydroxyalkanoates by genetically modified Methylobacterium extorquens strains.Microbial cell factories · 2010Article
- Phage Mu-driven two-plasmid system for integration of recombinant DNA in the Methylophilus methylotrophus genome.Applied microbiology and biotechnology · 2008Article
- Production of an insecticidal crystal protein from Bacillus thuringiensis by the methylotroph Methylobacterium extorquens.Applied and environmental microbiology · 2008Article
- Bestowing inducibility on the cloned methanol dehydrogenase promoter (PmxaF) of Methylobacterium extorquens by applying regulatory elements of Pseudomonas putida F1.Applied and environmental microbiology · 2006Article
- Fast, easy and efficient: site-specific insertion of transgenes into enterobacterial chromosomes using Tn7 without need for selection of the insertion event.BMC microbiology · 2006Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
High-level expression of chromosomally integrated genes in Methylobacterium extorquens ATCC 55366 was achieved under the control of the strong M. extorquens AM1 methanol dehydrogenase promoter (PmxaF) using the mini-Tn7 transposon system. Stable maintenance and expression of the integrated genes were obtained in the absence of antibiotic selective pressure. Furthermore, using this technology, a multicopy integration protocol for M. extorquens was also developed. Chromosomal integration of one to five copies of the gene encoding the green fluorescent protein (gfp) was achieved. The multicopy-based expression system permitted expression of a preset number of gene copies. A unique specific Tn7 integration locus in the chromosome of M. extorquens, known as the Tn7 attachment site (attTn7 site), was identified. This single attTn7 site was identified in an intergenic region between glmS, which encodes the essential enzyme glucosamine-6-phosphate synthetase, and dhaT, which encodes 1,3-propanediol dehydrogenase. The fact that the integration event is site specific and the fact that the attTn7 site is a noncoding region of the chromosome make the mini-Tn7 transposon system very useful for insertion of target genes and subsequent expression. In all transformants tested, expression and segregation of the transforming gene were stable without generation of secondary mutations in the host. In this paper, we describe single and multicopy chromosome integration and stable expression of heterologous genes (bgl [beta-galactosidase], est [esterase], and gfp [green fluorescent protein]) in M. extorquens.
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