Gross Chromosomal Rearrangements in Kluyveromyces marxianus Revealed by Illumina and Oxford Nanopore Sequencing.

Autor: Ding L; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA., Macdonald HD; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA., Smith HO; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA.; Codex DNA, 9535 Waples St #100, San Diego, CA 92121, USA., Hutchison Iii CA; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA., Merryman C; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA., Michael TP; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA., Abramson BW; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA., Kannan K; Codex DNA, 9535 Waples St #100, San Diego, CA 92121, USA., Liang J; Codex DNA, 9535 Waples St #100, San Diego, CA 92121, USA., Gill J; Codex DNA, 9535 Waples St #100, San Diego, CA 92121, USA., Gibson DG; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA.; Codex DNA, 9535 Waples St #100, San Diego, CA 92121, USA., Glass JI; J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA.
Jazyk: angličtina
Zdroj: International journal of molecular sciences [Int J Mol Sci] 2020 Sep 26; Vol. 21 (19). Date of Electronic Publication: 2020 Sep 26.
DOI: 10.3390/ijms21197112
Abstrakt: Kluyveromyces marxianus ( K. marxianus ) is an increasingly popular industrially relevant yeast. It is known to possess a highly efficient non-homologous end joining (NHEJ) pathway that promotes random integration of non-homologous DNA fragments into its genome. The nature of the integration events was traditionally analyzed by Southern blot hybridization. However, the precise DNA sequence at the insertion sites were not fully explored. We transformed a PCR product of the Saccharomyces cerevisiae URA3 gene ( ScURA3 ) into an uracil auxotroph K. marxianus otherwise wildtype strain and picked 24 stable Ura+ transformants for sequencing analysis. We took advantage of rapid advances in DNA sequencing technologies and developed a method using a combination of Illumina MiSeq and Oxford Nanopore sequencing. This approach enables us to uncover the gross chromosomal rearrangements (GCRs) that are associated with the ScURA3 random integration. Moreover, it will shine a light on understanding DNA repair mechanisms in eukaryotes, which could potentially provide insights for cancer research.
Competing Interests: The authors declare no conflict of interest.
Databáze: MEDLINE
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