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. 2016 Feb 1;36(4):628–644. doi: 10.1128/MCB.00436-15

TABLE 5.

The half-life of ADH2 mRNA made in glucose-containing medium is affected by the activatora

Plasmid Activator Half-life (min)b Mean relative abundance (SD) of ADH2 mRNA atc:
25°C 36.5°C
pKD16 Adr1 65 0.74 (0.36) 0.33 (0.13)
pKD17 oeAdr1 56 21 (7.5) 10 (8.3)
pAG11 Adr1-Gal11 10 28 (19) 0.5 (0.14)
a

Strain KBY108 (relevant genotype, rpb1-1 trp1) was transformed with the TRP1-containing plasmids pKD16, pKD17, and pAG11. Three transformants of each type were grown in synthetic complete (SC) medium lacking tryptophan and containing 5% glucose at 25°C to an A600 of 0.8. Samples were taken for RNA isolation and analysis, and the temperature of the cultures was raised to 36.5°C by adding an equal volume of 50°C medium. After 60 min of vigorous shaking, the cultures were collected for RNA isolation and analysis as described in Materials and Methods.

b

The apparent half-life of ADH2 mRNA was estimated as described in the legend to Table 3 from a graph of log(ADH2 mRNA/18S rRNA) versus time using the 0- and 60-min time points.

c

The abundance of ADH2 mRNA and 18s rRNA was determined by RT-qPCR as described in Materials and Methods. Each value represents the mean ADH2/18S RNA level (multiplied by 1,000) and standard deviation from three replicate cultures.