Using the structural gene for the ribosomal protein L3 from Saccharomyces cerevisiae as a probe, we isolated a homologous fragment from genomic DNA of Schizosaccharomyces pombe. Analysis of the plasmid carrying this fragment by hybridization selection and 2D-electrophoresis revealed a 31 kDa ribosomal protein. Transformation of the vector pDB248x containing this fragment into Schizosaccharomyces pombe leads to an increased level of mRNA suggesting that we have cloned the entire and actively transcribed gene.
To define the extent of intervening sequences required for efficient splicing of the CYH2 gene in Saccharomyces cerevisiae, we have constructed a series of intron mutations. Artificial intron extensions of more than 300 bp of the natural intron lead to an inhibition of splicing whereas intron deletions lead to a drastic improvement of the splicing efficiency. It is shown that deletion of a 32 bp sequence element within the intron is responsible for this drastic improvement.
We have developed an efficient transcriptlon system in isolated yeast nuclei. If MnC12 is substituted by CdC12, degradation of newly synthesrzed RNA is markedly reduced. This effect is due to the inhibition of nuclear ribonuclease activity, since microsomal ribonuclease activity is less affected by the cation, The extent to which the addition of CdCL to the in vitro transcription assay inhibits ribonuclease activity is demonstrated by the measurements of the size of newly synthesized RNA. Efficient RNA synthesis in this system is not affected up to a concentration of 0.1 M CdCl2.Yeast Cadmium RNA synthesis Rlbonuclease
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