Archaeal pyrrolysyl tRNA synthetases for orthogonal use
AssigneeEuropean Molecular Biology Laboratory
InventorsEdward Lemke, Ivana Nikic, Gemma Estrada Girona, Christine Köhler
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Abstract
The invention relates to archaeal pyrrolysyl tRNA synthetases lacking a nuclear localization signal and/or comprising a nuclear export signal. The invention also relates to polynucleotides encoding said pyrrolysyl tRNA synthetases, eukaryotic cells comprising said polynucleotide and tRNA acylated by the pyrrolysyl tRNA synthetase or a polynucleotide encoding such tRNA, methods utilizing said cells for preparing polypeptides comprising unnatural amino acid residues, and kits useful in said methods.
Claims11
A polynucleotide encoding a modified Methanosarcina mazei pyrrolysyl tRNA synthetase sequence or a Methanosarcina barkeri pyrrolysyl tRNA synthetase sequence wherein the modified pyrrolysyl tRNA synthetases are modified by introduction of a nuclear export signal, while retaining pyrrolysyl tRNA synthetase activity;
- wherein said pyrrolysyl tRNA synthetase comprises a Methanosarcina pyrrolysyl tRNA synthetase comprising a Methanosarcina mazei pyrrolysyl tRNA synthetase of SEQ ID NO:
- 1 or 2, a Methanosarcina barkeri pyrrolysyl tRNA synthetase of SEQ ID NO:
- 3, or a sequence having at least 90% sequence identity to any one of the sequences of SEQ ID NO:
- 1, 2 or 3.
Dependencies are inferred from the claim text.