Nanopore sequencing enables near-complete de novo assembly of Saccharomyces cerevisiae reference strain CEN.PK113-7D

Nanopore sequencing enables near-complete de novo assembly of Saccharomyces cerevisiae reference strain CEN.PK113-7D

Salazar, A.N. (TU Delft Pattern Recognition and Bioinformatics; Broad Institute of MIT and Harvard)
Gorter de Vries, A.R. (TU Delft BT/Industrial Microbiology)
van den Broek, M.A. (TU Delft BT/Industrial Microbiology)
Wijsman, M. (TU Delft BT/Industrial Microbiology)
de la Torre, P. (TU Delft BT/Industrial Microbiology)
Brickwedde, A. (TU Delft BT/Industrial Microbiology)
Brouwers, N. (TU Delft BT/Industrial Microbiology)
Daran, J.G. (TU Delft BT/Industrial Microbiology)
Abeel, T.E.P.M.F. (TU Delft Pattern Recognition and Bioinformatics; Broad Institute of MIT and Harvard)

2017

The haploid Saccharomyces cerevisiae strain CEN.PK113–7D is a popular model system for metabolic engineering and systems biology research. Current genome assemblies are based on short-read sequencing data scaffolded based on homology to strain S288C. However, these assemblies contain large sequence gaps, particularly in subtelomeric regions, and the assumption of perfect homology to S288C for scaffolding introduces bias. In this study, we obtained a near-complete genome assembly of CEN.PK113–7D using only Oxford Nanopore Technology's MinION sequencing platform. Fifteen of the 16 chromosomes, the mitochondrial genome and the 2-μm plasmid are assembled in single contigs and all but one chromosome starts or ends in a telomere repeat. This improved genome assembly contains 770 Kbp of added sequence containing 248 gene annotations in comparison to the previous assembly of CEN.PK113–7D. Many of these genes encode functions determining fitness in specific growth conditions and are therefore highly relevant for various industrial applications. Furthermore, we discovered a translocation between chromosomes III and VIII that caused misidentification of a MAL locus in the previous CEN.PK113–7D assembly. This study demonstrates the power of long-read sequencing by providing a high-quality reference assembly and annotation of CEN.PK113–7D and places a caveat on assumed genome stability of microorganisms.

Saccharomyces cerevisiae
yeast
genome assembly
long read sequencing
nanopore sequencing

http://resolver.tudelft.nl/uuid:7eaaad1f-e886-426b-bc72-4ec940eea742

https://doi.org/10.1093/femsyr/fox074

1567-1364

FEMS Yeast Research, 17 (7), 1-11

Institutional Repository

journal article

© 2017 A.N. Salazar, A.R. Gorter de Vries, M.A. van den Broek, M. Wijsman, P. de la Torre, A. Brickwedde, N. Brouwers, J.G. Daran, T.E.P.M.F. Abeel