T7 rna polymerase variants for rna synthesis
AssigneeGlaxosmithkline Biologicals SA
InventorsBrent M. DORR, Joel MELBY, Deepak RUDRAPPA, Jeliazko JELIAZKOV, Nilesh VAIDYA
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Abstract
The present disclosure provides T7 RNA polymerase variants, the use of which increases transcription efficiency and improves the quality and yield of the enzyme. Also provided are polynucleotides encoding the T7 RNA polymerase variants, host cells capable of expressing the T7 RNA polymerase variants, and methods of using the T7 RNA polymerase variants for high-efficiency transcription.
Claims30
A T7 ribonucleic acid (RNA) polymerase comprising an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence set out in SEQ ID NO:1 or a functional fragment thereof, wherein the T7 RNA polymerase amino acid sequence includes the feature that:
- at least one amino acid selected from the group consisting of amino acid N15, I20, Q64, E69, A257, A264, A268, T381, N425, D427, M437, E649, Q655, D659, G681, W688, F761, K779, and I784 is substituted to a different amino acid to that found at the corresponding amino acid position in SEQ ID NO:1.
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A T7 RNA polymerase comprising an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence set out in SEQ ID NO:2 or a functional fragment thereof, wherein the T7 RNA polymerase amino acid sequence includes the feature that:
- at least one amino acid selected from the group consisting of amino acid N8, I13, Q57, E62, A250, A257, A261, T374, N418, D420, M430, E642, Q648, D652, G674, W681, F754, K772, and I777 is substituted to a different amino acid to that found at the corresponding position in SEQ ID NO:
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