ArticleScientific reports2015
A rapid and reliable strategy for chromosomal integration of gene(s) with multiple copies.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers, 1 of them a synthesis that pooled it.
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Who cites it
28 citing papers in PubMed, 1 synthesis or guideline pooled it, 77 citations in OpenAlex.
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- Advances and prospects in metabolic engineering of Escherichia coli for L-tryptophan production.World journal of microbiology & biotechnology · 2022Review
- Optimising PHBV biopolymer production in haloarchaea via CRISPRi-mediated redirection of carbon flux.Communications biology · 2021Article
- Expansion of EasyClone-MarkerFree toolkit for Saccharomyces cerevisiae genome with new integration sites.FEMS yeast research · 2021Article
- Construction of recombinant Escherichia coli for production of L-phenylalanine-derived compounds.World journal of microbiology & biotechnology · 2021Review
- Precision design of stable genetic circuits carried in highly-insulated E. coli genomic landing pads.Molecular systems biology · 2020Article
- Enhanced succinic acid production by Mannheimia employing optimal malate dehydrogenase.Nature communications · 2020Article
- Targeted and Repetitive Chromosomal Integration Enables High-Level Heterologous Gene Expression inApplied and environmental microbiology · 2019Article
- Advanced Strategies for Food-Grade Protein Production: A NewMicroorganisms · 2019Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Direct optimization of the metabolic pathways on the chromosome requires tools that can fine tune the overexpression of a desired gene or optimize the combination of multiple genes. Although plasmid-dependent overexpression has been used for this task, fundamental issues concerning its genetic stability and operational repeatability have not been addressed. Here, we describe a rapid and reliable strategy for chromosomal integration of gene(s) with multiple copies (CIGMC), which uses the flippase from the yeast 2-μm plasmid. Using green fluorescence protein as a model, we verified that the fluorescent intensity was in accordance with the integration copy number of the target gene. When a narrow-host-range replicon, R6K, was used in the integrative plasmid, the maximum integrated copy number of Escherichia coli reached 15. Applying the CIGMC method to optimize the overexpression of single or multiple genes in amino acid biosynthesis, we successfully improved the product yield and stability of the production. As a flexible strategy, CIGMC can be used in various microorganisms other than E. coli.
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