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AssigneeYale University
InventorsDieter Soll, Caroline Aldag, Michael Hohn, Corwin Miller
Estimating the term…
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.
An isolated nucleic acid comprising a nucleic acid sequence encoding a non-naturally occurring tRNASec, wherein the non-naturally occurring tRNASec is recognized by SerRS, when aminoacylated with serine the Ser-tRNASec is a substrate for SelA, and when aminoacylated with seleocysteine the Sec-tRNASec is recognized by EF-Tu,
A non-naturally occurring tRNAsec comprising a sequence at least 80% identical to SEQ ID NO:6, 7, or 8, wherein the tRNAsec is recognized by SerRS, when aminoacylated with serine the Ser-tRNASec is a substrate for SelA, and when aminoacylated with seleocysteine the Sec-tRNASec is recognized by EF-Tu.
A non-naturally occurring tRNAsec comprising the nucleic acid sequence of any one of SEQ ID NO:59-85, or a variant thereof with a substituted anticodon.
Dependencies are inferred from the claim text.