Effects of ABO intron 1 on transcriptional activation when the +5653 to +6154 sequence was inserted upstream of various promoters
| Reporter construct . | Promoter . | Insert . | Relative activity† . | -fold activity over control . |
|---|---|---|---|---|
| Test plasmids | ||||
| C/SN | ABO proximal promoter | +5653: +6154‡ | 2.386 ± 0.170 | 2.4 |
| C/TK | TK promoter | +5653: +6154‡ | 8.041 ± 1.583 | 2.0 |
| C/SV | SV40 promoter | +5653: +6154‡ | 7.247 ± 0.170 | 1.3 |
| SN/C | ABO proximal promoter | +5653: +6154§ | 1.497 ± 0.140 | 1.5 |
| Controls | ||||
| SN | ABO proximal promoter | None | 1.000 | |
| TK-luc | TK promoter | None | 4.023 ± 0.718 | |
| pGL3-promoter | SV40 promoter | None | 5.555 ± 1.315 | |
| pGL3-basic | None | None | 0.060 ± 0.014 | |
| Reporter construct . | Promoter . | Insert . | Relative activity† . | -fold activity over control . |
|---|---|---|---|---|
| Test plasmids | ||||
| C/SN | ABO proximal promoter | +5653: +6154‡ | 2.386 ± 0.170 | 2.4 |
| C/TK | TK promoter | +5653: +6154‡ | 8.041 ± 1.583 | 2.0 |
| C/SV | SV40 promoter | +5653: +6154‡ | 7.247 ± 0.170 | 1.3 |
| SN/C | ABO proximal promoter | +5653: +6154§ | 1.497 ± 0.140 | 1.5 |
| Controls | ||||
| SN | ABO proximal promoter | None | 1.000 | |
| TK-luc | TK promoter | None | 4.023 ± 0.718 | |
| pGL3-promoter | SV40 promoter | None | 5.555 ± 1.315 | |
| pGL3-basic | None | None | 0.060 ± 0.014 | |
The constructs as depicted above were transiently transfected into K562 cells; 2.5 μg of luciferase reporter and 0.01 μg of Renilla luciferase control vector were used for each analysis.
The results are expressed as the average relative activity compared with that observed for reporter vector SN containing the ABO proximal promoter. SDs are indicated for a minimum of 3 repetitions.
The +5653 to +6154 sequence was subcloned upstream of the ABO proximal promoter, TK promoter, and SV40 promoter in an orientation opposite to the transcription direction in these promoters.
The +5653 to +6154 sequence was subcloned downstream of the luciferase gene in the same orientation as the transcription direction in the promoter.