1991
DOI: 10.1016/0076-6879(91)94032-8
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[29] Measurement of mRNA decay rates in Saccharomyces cerevisiae

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Cited by 113 publications
(98 citation statements)
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“…Routinely, 9 cells were frozen within 15 sec of removal of the culture aliquot. Total yeast RNA was isolated as described previously Parker et al 1991). Equal amounts (usually 10-20 ~g) of total RNA from each time point of an experiment were analyzed by Northern blotting (Thomas 1980).…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Routinely, 9 cells were frozen within 15 sec of removal of the culture aliquot. Total yeast RNA was isolated as described previously Parker et al 1991). Equal amounts (usually 10-20 ~g) of total RNA from each time point of an experiment were analyzed by Northern blotting (Thomas 1980).…”
Section: Methodsmentioning
confidence: 99%
“…The numbers at left correspond to the various PGK1 alleles represented in B: (1) the wild-type PGK1 gene; (2) the PCK1 gene with a nonsense mutation at 92.6% of the coding region (inserted at the BgIII site; 385 PGK1 codons translated); {3) the PGKI gene with a nonsense mutation at 76.2% of the coding region (inserted at the XbaI site; 317 PGK1 codons translated); (4) the PGK1 gene with a nonsense mutation at 67.7% of the coding region [inserted at the H2(1) site; 282 PGK1 codons translated[; [5) the PGK1 gene with a nonsense mutation at 55% of the coding region [inserted at the H2{2) site; 229 PGK1 codons translated]; (6) the PGK1 gene with a nonsense mutation at 39% of the coding region (inserted at the Asp718 site; 164 PGK1 codons translated); (7) the PGK1 gene with a nonsense mutation at 5.6% of the coding region [inserted at the H2{3) site; 23 PGK1 codons translated]. The decay assay, RNA blotting, hybridization, and quantitation of the blots were performed as described previously Parker et al 1991). (B) mRNA abundance and decay rates for the wild-type and mutant PGK1 alleles in SWP154( + ) (wild-type for the UPF1 gene) or SWP154( -) (UPFI(-) (a deletion of the UPF1 gene), mRNA abundance was measured by RNA blot analysis of equal amounts of RNA from the time zero points from each of the mRNA decay measurements, normalizing to the abundance of the CYH2 transcript.…”
Section: Destabilization Of the Pgk1 Transcript Is Dependent On The Pmentioning
confidence: 99%
“…The strains to be tested were grown overnight at 23°C to log phase. Cell cultures were concentrated fivefold and then shifted to 37°C (43). At the indicated time points 2-ml aliquots of the cultures were quickly collected and frozen in dry ice-ethanol.…”
Section: Strainsmentioning
confidence: 99%
“…To determine whether the differences we observed are due to a difference in the splicing rate, we measured the half-lives of the spliced intron RNA and each pre-rRNA after inhibition of GAL transcription by glucose at 30°C (Parker et al 1991). Addition of glucose inhibits the expression of GAL genes within 1-2 min (Ronen and Botstein 2006).…”
Section: Rrna Improves Fidelity Of Foldingmentioning
confidence: 99%
“…The cells were harvested and resuspended in 15 mL of minimal medium with 2% glucose at 30°C to prevent further transcription from the GAL promoter (Parker et al 1991). Aliquots (1 mL) were harvested at various times by spinning in a microcentrifuge for 10 sec at 14,000g.…”
Section: Rna Decay Ratesmentioning
confidence: 99%