TABLE 5.

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

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)

^aStrain 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.

^bThe 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.

^cThe 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.