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PMID: 3892482 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Evaluation of heterologous ARS activity in S. cerevisiae using cloned DNA from S. pombe.

Nucleic acids research ·Vol. 13 ·No. 10 ·1985-05-24 ·Pages 3711-22

Maundrell K, Wright AP, Piper M, Shall S

Abstract

Cloned segments of Schizosaccharomyces pombe genomic DNA were screened for ARS activity in the native host, S. pombe, using high frequency transformation, phenotypic instability and extrachromosomal maintenance of unrearranged plasmid sequences as criteria for ARS function. This analysis revealed 12 ARS elements in a total of 230 kb of chromosomal DNA, indicating an average frequency of one ARS every 19 kb of genomic DNA. We then used these clones to assess the reliability of the S. cerevisiae assay for detecting ARS elements in heterologous DNA. The results show that not only does the S. cerevisiae assay fail to detect a large proportion of true ARS elements but it also wrongly identifies a significant proportion of clones which did not display ARS activity in the native host. We would therefore recommend restraint when extrapolating from observed ARS function of heterologous DNA in S. cerevisiae to a presumed analogous role in the original host.

MeSH Terms
Base Sequence Cloning, Molecular DNA Replication DNA, Fungal/metabolism Saccharomyces/genetics Saccharomyces cerevisiae/genetics Transformation, Genetic
Chemicals
DNA, Fungal
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Maundrell K
Wright A P
Piper M
Shall S
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22 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.10.3711
Published
1985-05-24
Pages
3711-22
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341268
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/13.10.3711 PMC Full Text View journal details
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