TABLE 7.4.
Experiment to show that expression of a subgenomic RNA (sGFP) from a construct in which the coat protein gene of RCNMV RNA1 is replaced by GFP (R1sGFP) depends on RNA2 sequences
| R1sGFP was co-inoculated with transcripts from the TBSV replicon (pHST2 in which the coat protein region was engineered to accept and express foreign genes from an sgRNA) containing segments from RCNMV RNA2 in order to delimit the minimal trans-activating elements. | |||
|---|---|---|---|
| Construct | Position of RNA2 sequence | Length of RNA2 inserts (nucleotides) | sGFP produceda |
| RCNMV RNA2 | 1–448 | 1448 | + + + |
| pHST2 | − | − | − |
| pHST2-RC2.3 | 708–1031 | 324 | + |
| pHST2-RC2.4 | 1031–708 (-sense) | 324 | − |
| pHST2-ΔBX | 708–916 | 209 | + + |
| pHST2–707 | 707–837 | 121 | + |
| pHST2–828 | 828–918 | 91 | − |
| pHST2-SL2 | 756–789 | 34 | + + + |
| pHST2-TA38 | 792–755 (-sense) | 38 | − |
| pHST2–20 | 762–782 | 21 | + |
| pHST2-LT2 | 767–775 | 9 | − |
a
sGFP production relative to induction by RNA2: + + + = 100%; + + = 50%; + = 25%; – = not detected. From Sit et al. (1998), with permission.