Key result
The presence of one extra base pair G.C upstream of the T7 promoter increased transcription by one order of magnitude compared to a canonical T7 promoter sequence.
The addition of a single G.C base pair upstream of the T7 promoter significantly enhances T7 RNA polymerase-dependent transcription in vitro.
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May enhance in vitro transcription tools; leaves open validation in cardiovascular gene expression models.
The T7 RNA polymerase-dependent transcription was studied as a function of nucleotide sequence structures positioned upstream of the T7 promoter. Model double-stranded DNA templates were constructed for this purpose. They contained a target sequence of 485 base pairs (cDNA fragment of Venesuelian encephalomyelitis equine virus genome), T7 promoter consensus and different extra base sequences upstream of the T7 promoter. The level of the target sequence transcription was clearly determined by the extra base sequence. The presence of one extra base pair G.C ensured the most pronounced effect, transcription was increased one order of magnitude in comparison with template which has only a canonical T7 promoter sequence at the 5'-end.
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Michail M. Baklanov (1996) studied this question. Extra base sequences upstream of the T7 promoter (one extra base pair G.C) vs. Template with only a canonical T7 promoter sequence at the 5'-end was evaluated on Level of target sequence transcription (increased one order of magnitude). The presence of one extra base pair G.C upstream of the T7 promoter increased transcription by one order of magnitude compared to a canonical T7 promoter sequence.
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