Table 2.
Description of constructs and mapping of a cold-shock response sequence to the EP1 3′-UTR
| Construct | Genomic locus | Promoter | 5′-UTR | Reporter gene | 3′-UTR | Cold-shock induction factora |
|---|---|---|---|---|---|---|
| pL20-wt.EG | rDNA spacer | GPEETTI | GPEET | EP1:GFP | EP1 | 64.5 |
| pG-Δ164.EG | EP | EP1 | EP1 | EP1:GFP | EP1Δ1-164 | 62 |
| pG-ΔLII.EG | EP | EP1 | EP1 | EP1:GFP | EP1Δ101-173 | 61 |
| pG-ΔLII.Luc | EP | EP1 | EP1 | luciferase | EP1Δ101-173 | 65.1 |
| pL20-ΔLII.EG | rDNA spacer | GPEETTI | GPEET | EP1:GFP | EP1Δ101-173 | 63.7 |
| pL82-ΔLII.EG | rDNA spacer | T7TI | GPEET | EP1:GFP | EP1Δ101-173 | >30b |
| pTub-ΔLII.EG | tubulin | Read through | aldolase | EP1:GFP | EP1Δ101-173 | 59.8 |
| pL20-ΔUTR.EG | rDNA spacer | GPEETTI | GPEET | EP1:GFP | EP1Δ1-297 | 3.2 |
| pL20-ΔUTR.Luc | rDNA spacer | GPEETTI | GPEET | luciferase | EP1Δ1-297 | 5.5 |
| pES221-EG | vsg221 ES | VSG | actin | EP1:GFP | actin | 4.7 |
| pL82-EG | rDNA spacer | T7TI | GPEET | EP1:GFP | aldolase 1-177 | 5.9 |
| pL20-Luc | rDNA spacer | GPEETTI | GPEET | luciferase | aldolase 1-177 | 6.3 |
| pTR-EG | rDNA promoter | rDNA | EP1 | EP1-GFP | tubulin | 3.7 |
a
Quotient of reporter expression measured at 20°C and 37°C. A table with the underlying data is provided as Supplemental Material
b
Due to the high basal expression level, a cold-shock-induced elevation of EP1:GFP of >30-fold is lethal for the cells