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Protocol
Methods to Assess Phosphodiesterase and/or Adenylyl Cyclase Activity Via Heterologous Expression in Fission Yeast
Heterologous expression of cyclic nucleotide phosphodiesterases (PDEs) and adenylyl cyclases (ACs) in the fission yeast Schizosaccharomyces pombe can be used in combination with PKA-repressed reporters to either ...
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Article
Open AccesslncRNA transcription induces meiotic recombination through chromatin remodelling in fission yeast
Noncoding RNAs (ncRNAs) are involved in various biological processes, including gene expression, development, and disease. Here, we identify a novel consensus sequence of a cis-element involved in long ncRNA (lnc...
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Article
Open AccesslncRNA transcriptional initiation induces chromatin remodeling within a limited range in the fission yeast fbp1 promoter
Long noncoding RNAs (lncRNAs) transcribed across gene promoters have been detected. These regulate transcription by mechanisms that have not been fully elucidated. We herein show that the chromatin configurati...
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Protocol
A Yeast-Based High-Throughput Screen for Modulators of Phosphodiesterase Activity
Cell-based high-throughput screens (HTSs) targeting heterologously expressed proteins in yeast identify compounds that often display relevant biological activity when tested in cell culture. We developed a fis...
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Protocol
Fission Yeast-Based High-Throughput Screens for PKA Pathway Inhibitors and Activators
Features of the fission yeast Schizosaccharomyces pombe cAMP/PKA pathway make S. pombe particularly amenable for heterologous expression of cAMP pathway proteins such as GαS subunits and their cognate adenylyl cy...
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Article
Use of a ura5 +–lys7 + cassette to construct unmarked gene knock-ins in Schizosaccharomyces pombe
While the counterselectable Schizosaccharomyces pombe ura4 + gene can be used to prepare a site in the S. pombe genome to receive an unmarked mutant allele (loss of ura4 ...
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Chapter
A Fission Yeast-Based Platform for Phosphodiesterase Inhibitor HTSs and Analyses of Phosphodiesterase Activity
Fission yeast strains have been engineered so that their growth behavior reflects the activity of heterologous cyclic nucleotide phosphodiesterases (PDEs). These strains can be used in High-Throughput Screens ...
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Article
Stepwise chromatin remodelling by a cascade of transcription initiation of non-coding RNAs
Non-coding RNAs are abundant, yet their function is still largely unknown. Here, non-coding RNA transcripts are detected at the fission yeast fbp1+ locus, upstream of the normal transcription initiation site. ...
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Article
Cloning the Schizosaccharomyces pombe lys2 + gene and construction of new molecular genetic tools
Molecular genetic analyses in Schizosaccharomyces pombe rely on selectable markers that are used in cloning vectors or to mark targeted gene deletions and other integrated constructs. In this study, we used genet...
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Article
Cloning and manipulation of the Schizosaccharomyces pombe his7 +gene as a new selectable marker for molecular genetic studies
We have cloned the his7 + gene of the fission yeast Schizosaccharomyces pombe by complementation of the recessive mutant allele his7-366. The his7 + gene is able to complement a mutation of the Escherichia coli h...
