Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Version 1 : Received: 30 August 2020 / Approved: 31 August 2020 / Online: 31 August 2020 (02:54:09 CEST)

How to cite: Ding, L.; Macdonald, H.D.; Smith, H.O.; Hutchison III, C.A.; Merryman, C.; Michaels, T.P.; Abramson, B.W.; Kannan, K.; Liang, J.; Gill, J.; Gibson, D.G.; Glass, J.I. Gross Chromosomal Rearrangements in Kluyveromyces Marxianus Revealed by Illumina and Oxford Nanopore Sequencing. Preprints 2020, 2020080687 (doi: 10.20944/preprints202008.0687.v1). Ding, L.; Macdonald, H.D.; Smith, H.O.; Hutchison III, C.A.; Merryman, C.; Michaels, T.P.; Abramson, B.W.; Kannan, K.; Liang, J.; Gill, J.; Gibson, D.G.; Glass, J.I. Gross Chromosomal Rearrangements in Kluyveromyces Marxianus Revealed by Illumina and Oxford Nanopore Sequencing. Preprints 2020, 2020080687 (doi: 10.20944/preprints202008.0687.v1).

Abstract

Kluyveromyces marxianus (K. marxianus) is a newly emerging 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 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.

Subject Areas

gross chromosomal rearrangements; non-homologous end joining; translocation; Illumina MiSeq; Oxford Nanopore; kluyveromyces marxianus; saccharomyces cerevisiae; URA3 gene

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