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Abstract <p>To date, the genomes of several mycoplasmas have been cloned in <i>Saccharomyces cerevisiae</i> cells, including <i>Mycoplasma genitalium</i>, <i>M. pneumoniae</i>, and <i>M. mycoides</i>. In this work, the 1-Mb genome of <i>Mycoplasma gallisepticum</i> S6 was cloned in <i>S. cerevisiae</i>. A transposon vector was constructed to deliver a synthetic <i>S. cerevisiae</i> replicon and a selective marker into <i>M. gallisepticum</i> genome. Transplantation of the <i>M. gallisepticum</i> genome with the integrated yeast replicon to yeast cells was performed using two approaches, polyethylene glycol (PEG)-induced transformation of yeast spheroplasts with intact genomic DNA isolated from <i>M.&#xa0;gallisepticum</i> cells and prepared in aragose plugs and PEG-induced fusion of <i>M. gallisepticum</i> cells with yeast spheroplasts. Both of the approaches yielded <i>S. cerevisiae</i> strains with the cloned <i>M. gallisepticum</i> genome, as was verified by nanopore sequencing.</p>

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Cloning of Mycoplasma gallisepticum Genome in Saccharomyces cerevisiae Cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月2日

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  • D. R. Kharrasov,
  • G. Yu. Fisunov,
  • D. S. Matyushkina,
  • E. A. Tsoy,
  • E. A. Semenova,
  • R. R. Kutukov,
  • I. S. Rog,
  • D. A. Sandanova,
  • M. M. Borodina,
  • V. M. Govorun

摘要

Abstract

To date, the genomes of several mycoplasmas have been cloned in Saccharomyces cerevisiae cells, including Mycoplasma genitalium, M. pneumoniae, and M. mycoides. In this work, the 1-Mb genome of Mycoplasma gallisepticum S6 was cloned in S. cerevisiae. A transposon vector was constructed to deliver a synthetic S. cerevisiae replicon and a selective marker into M. gallisepticum genome. Transplantation of the M. gallisepticum genome with the integrated yeast replicon to yeast cells was performed using two approaches, polyethylene glycol (PEG)-induced transformation of yeast spheroplasts with intact genomic DNA isolated from M. gallisepticum cells and prepared in aragose plugs and PEG-induced fusion of M. gallisepticum cells with yeast spheroplasts. Both of the approaches yielded S. cerevisiae strains with the cloned M. gallisepticum genome, as was verified by nanopore sequencing.