Format

Send to

Choose Destination
See comment in PubMed Commons below
Genome Res. 2013 Apr;23(4):698-704. doi: 10.1101/gr.144659.112. Epub 2012 Dec 12.

High-resolution mapping, characterization, and optimization of autonomously replicating sequences in yeast.

Author information

1
Department of Genome Sciences, University of Washington, Seattle, Washington 98105, USA.

Abstract

DNA replication origins are necessary for the duplication of genomes. In addition, plasmid-based expression systems require DNA replication origins to maintain plasmids efficiently. The yeast autonomously replicating sequence (ARS) assay has been a valuable tool in dissecting replication origin structure and function. However, the dearth of information on origins in diverse yeasts limits the availability of efficient replication origin modules to only a handful of species and restricts our understanding of origin function and evolution. To enable rapid study of origins, we have developed a sequencing-based suite of methods for comprehensively mapping and characterizing ARSs within a yeast genome. Our approach finely maps genomic inserts capable of supporting plasmid replication and uses massively parallel deep mutational scanning to define molecular determinants of ARS function with single-nucleotide resolution. In addition to providing unprecedented detail into origin structure, our data have allowed us to design short, synthetic DNA sequences that retain maximal ARS function. These methods can be readily applied to understand and modulate ARS function in diverse systems.

PMID:
23241746
PMCID:
PMC3613586
DOI:
10.1101/gr.144659.112
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for HighWire Icon for PubMed Central
    Loading ...
    Support Center