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

Ding, Lin ; Macdonald, Harrison D ; Smith, Hamilton O ; Hutchison Iii, Clyde A ; Merryman, Chuck ; Michael, Todd P ; Abramson, Bradley W ; Kannan, Krishna ; Liang, Joe ; Gill, John ; Gibson, Daniel G ; Glass, John I

International journal of molecular sciences, 2020-10, Vol.21 (19), p.7112 [Periódico revisado por pares]

Switzerland: MDPI AG

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Título:
Gross Chromosomal Rearrangements in Kluyveromyces marxianus Revealed by Illumina and Oxford Nanopore Sequencing
Autor: Ding, Lin ; Macdonald, Harrison D ; Smith, Hamilton O ; Hutchison Iii, Clyde A ; Merryman, Chuck ; Michael, Todd P ; Abramson, Bradley W ; Kannan, Krishna ; Liang, Joe ; Gill, John ; Gibson, Daniel G ; Glass, John I
Assuntos: Ascomycota ; Cancer research ; Chromosome Aberrations ; Chromosome rearrangements ; Chromosomes ; Chromosomes, Fungal ; Cloning ; Deoxyribonucleic acid ; DNA ; DNA End-Joining Repair ; DNA repair ; DNA sequencing ; DNA, Fungal - genetics ; Eukaryotes ; Fungi ; Genetic aspects ; Genetic engineering ; Genomes ; gross chromosomal rearrangements ; Homology ; Hybridization ; Illumina MiSeq ; Integration ; Kluyveromyces - genetics ; Kluyveromyces marxianus ; Methods ; Nanopore Sequencing - methods ; Non-homologous end joining ; Nucleotide sequence ; Nucleotide sequencing ; Oxford Nanopore ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae Proteins - genetics ; Sequence analysis ; Transformation, Genetic ; translocation ; URA3 gene ; Uracil ; Yeast
É parte de: International journal of molecular sciences, 2020-10, Vol.21 (19), p.7112
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These authors contributed equally to this work.
Descrição: ( ) 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 gene ( ) into an uracil auxotroph 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 random integration. Moreover, it will shine a light on understanding DNA repair mechanisms in eukaryotes, which could potentially provide insights for cancer research.
Editor: Switzerland: MDPI AG
Idioma: Inglês

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

Ding, Lin ; Macdonald, Harrison D ; Smith, Hamilton O ; Hutchison Iii, Clyde A ; Merryman, Chuck ; Michael, Todd P ; Abramson, Bradley W ; Kannan, Krishna ; Liang, Joe ; Gill, John ; Gibson, Daniel G ; Glass, John I

Switzerland: MDPI AG

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