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• W4308915287 abstract "Abstract Background Natural life systems can be significantly modified at the genomic scale by human intervention, demonstrating the great innovation capacity of genome engineering. Large epi-chromosomal DNA structures were established in Escherichia coli cells, but some of these methods were inconvenient, using heterologous systems, or relied on engineered E. coli strains. Results The wild-type model bacterium E. coli has a single circular chromosome. In this work, a novel method was developed to split the original chromosome of wild-type E. coli . With this method, novel E. coli strains containing two chromosomes of 0.10 Mb and 4.54 Mb, and 2.28 Mb and 2.36 Mb were created respectively, designated as E. coli 0.10/4.54 and E. coli 2.28/2.36 . The new chromosomal arrangement was proved by PCR amplification of joint regions as well as a combination of Nanopore and Illumina sequencing analysis. While E. coli 0.10/4.54 was quite stable, the two chromosomes of E. coli 2.28/2.36 population recombined into a new chromosome (Chr.4.64M Mut ), via recombination. Both engineered strains grew slightly slower than the wild-type, and their cell shapes were obviously elongated. Conclusion Finally, we successfully developed a simple CRISPR-based genome engineering technique for the construction of multi-chromosomal E. coli strains with no heterologous genetic parts. This technique might be applied to other prokaryotes for synthetic biology studies and applications in the future." @default.
• W4308915287 created "2022-11-19" @default.
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• W4308915287 date "2022-11-11" @default.
• W4308915287 modified "2023-10-16" @default.
• W4308915287 title "A CRISPR-based chromosomal-separation technique for Escherichia coli" @default.
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• W4308915287 doi "https://doi.org/10.1186/s12934-022-01957-4" @default.
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