Evaluation of heterologous ARS activity in S. cerevisiae using cloned DNA from S. pombe.
AUTOR(ES)
Maundrell, K
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=341268Documentos Relacionados
- Genetic analysis of an ARS element from the fission yeast Schizosaccharomyces pombe.
- Identification and characterization of a telomerase activity from Schizosaccharomyces pombe.
- RNase-sensitive DNA modification(s) initiates S. pombe mating-type switching
- The 5S RNA genes of Schizosaccharomyces pombe.
- A single-stranded DNA binding protein from S. cerevisiae specifically recognizes the T-rich strand of the core sequence of ARS elements and discriminates against mutant sequences.