ArticleBMC molecular biology2009
Use of the lambda Red-recombineering method for genetic engineering of Pantoea ananatis.
Article in BMC molecular biology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed, 82 citations in OpenAlex.
- Glutamate dehydrogenase from Pantoea ananatis: A new bacterial enzyme with dual coenzyme specificity.PloS one · 2025Article
- Biosynthesis of C4-C8 3-Hydroxycarboxylic Acids from Glucose through the Inverted Fatty Acid β-Oxidation by Metabolically EngineeredBiomolecules · 2024Article
- Isolation and draft genome sequence ofJournal of genomics · 2024Article
- The emerging role of recombineering in microbiology.Engineering microbiology · 2023Review
- Recombineering in Non-Model Bacteria.Current protocols · 2022Article
- Article
- Revealing a New Family of D-2-Hydroxyglutarate Dehydrogenases inMicroorganisms · 2022Article
- Robust counterselection and advanced λRed recombineering enable markerless chromosomal integration of large heterologous constructs.Nucleic acids research · 2022Article
- Microbial Production Potential ofMicroorganisms · 2022Review
- TheMicroorganisms · 2022Article
- Fermentative production of enantiopure (S)-linalool using a metabolically engineered Pantoea ananatis.Microbial cell factories · 2021Article
- Ethylene glycol and glycolic acid production from xylonic acid by Enterobacter cloacae.Microbial cell factories · 2020Article
- Bacterial Genetic Engineering by Means of Recombineering for Reverse Genetics.Frontiers in microbiology · 2020Review
- Functional Characterization of a Global Virulence Regulator Hfq and Identification of Hfq-Dependent sRNAs in the Plant PathogenFrontiers in microbiology · 2019Article
- Article
- Mu-driven transposition of recombinant mini-Mu unit DNA in the Corynebacterium glutamicum chromosome.Applied microbiology and biotechnology · 2018Article
- Engineered Microorganisms for the Production of Food Additives Approved by the European Union-A Systematic Analysis.Frontiers in microbiology · 2018Review
- Swimming and twitching motility are essential for attachment and virulence of Pantoea ananatis in onion seedlings.Molecular plant pathology · 2017Article
- Fermentative Production of Cysteine by Pantoea ananatis.Applied and environmental microbiology · 2017Article
- Identification of enzymes responsible for extracellular alginate depolymerization and alginate metabolism in Vibrio algivorus.Applied microbiology and biotechnology · 2017Article
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Authors and funding
6 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundPantoea ananatis, a member of the Enterobacteriacea family, is a new and promising subject for biotechnological research. Over recent years, impressive progress in its application to L-glutamate production has been achieved. Nevertheless, genetic and biotechnological studies of Pantoea ananatis have been impeded because of the absence of genetic tools for rapid construction of direct mutations in this bacterium. The lambda Red-recombineering technique previously developed in E. coli and used for gene inactivation in several other bacteria is a high-performance tool for rapid construction of precise genome modifications.
resultsIn this study, the expression of lambda Red genes in P. ananatis was found to be highly toxic. A screening was performed to select mutants of P. ananatis that were resistant to the toxic affects of lambda Red. A mutant strain, SC17(0) was identified that grew well under conditions of simultaneous expression of lambda gam, bet, and exo genes. Using this strain, procedures for fast introduction of multiple rearrangements to the Pantoea ananatis genome based on the lambda Red-dependent integration of the PCR-generated DNA fragments with as short as 40 bp flanking homologies have been demonstrated.
conclusionThe lambda Red-recombineering technology was successfully used for rapid generation of chromosomal modifications in the specially selected P. ananatis recipient strain. The procedure of electro-transformation with chromosomal DNA has been developed for transfer of the marked mutation between different P. ananatis strains. Combination of these techniques with lambda Int/Xis-dependent excision of selective markers significantly accelerates basic research and construction of producing strains.
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