Compositions and methods for making selenocysteine containing polypeptides
AssigneeYale University
InventorsDieter Soll, Takahito Mukai, Kyle Hoffman
Life of this patent
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Abstract
Non-naturally occurring tRNASec and methods of using them for recombinant expression of proteins engineered to include one or more selenocysteine residues are disclosed. The non-naturally occurring tRNASec can be used for recombinant manufacture of selenocysteine containing polypeptides encoded by mRNA without the requirement of an SECIS element. In some embodiments, selenocysteine containing polypeptides are manufactured by co-expressing a non-naturally occurring tRNASec a recombinant expression system, such as E. coli, with SerRS, EF-Tu, SelA, or PSTK and SepSecS, and an mRNA with at least one codon that recognizes the anticodon of the non-naturally occurring tRNASec.
Claims19
A non-naturally occurring tRNASec comprising sequence elements from a yeast tRNASer in combination with sequence elements from a non-yeast tRNA,
- wherein the non-yeast tRNA is an Aeromonas salmonicida tRNA,
- wherein the sequence of the non-naturally occurring tRNASec comprises at least 85% sequence identity to the chimeric sequences of any of SEQ ID NO:
- 152, 154, 156 or SEQ ID NO:
- 158, and
- wherein the non-naturally occurring tRNASec is a substrate for a SerRS and a SelA, and binds to eEF1α.
A non-naturally occurring tRNASec comprising sequence elements from a yeast tRNASer in combination with sequence elements from a non-yeast tRNA,
- wherein the non-yeast tRNA is an Aeromonas salmonicida tRNA,
- wherein the sequence of the non-naturally occurring tRNASec comprises at least 90% sequence identity to the chimeric sequences of any of SEQ ID NO:
- 152, 154, 156 or SEQ ID NO:
- 158, and
- wherein the non-naturally occurring tRNASec is a substrate for a SerRS and a SelA and binds to eEF1α.
a non-naturally occurring tRNASec comprising any one of SEQ ID NO:152, 154, 156, or 158 or the opal or ochre equivalent thereof.
Dependencies are inferred from the claim text.