Method of combinatorial protein synthesis based on ribosomal frameshifting
OwnerWlodek Mandeckiinventor
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Abstract
Disclosed are materials and methods for practicing combinatorial protein synthesis based on ribosomal frameshifting. The genes, encoding the proteins to be synthesized, are constructed by assembly of several double-stranded DNA fragments and are cloned into bacteria. The genes include interspersed frameshifting sequences. Proteins are made in the cell by translating nucleotide sequences in a combinatorial mode. The invention can be used for the selection of new, modified or improved proteins.
Claims21
A method of producing a plurality of polypeptides from a single polynucleotide operably linked to a heterologous promoter comprising the step of expressing a single polynucleotide, wherein such polynucleotide comprises at least two ribosomal frameshifting sequences such that said single polynucleotide is capable of producing a plurality of different polypeptides.
The method of clai 1, wherein said ribosomal frameshifting sequence is a switch sequence.
A method of producing a plurality of peptides from a single polynucleotide operably linked to a heterologous promoter comprising the steps of:
- expressing a polynucleotide, wherein such polynucleotide comprises at least two ribosomal frameshifting sequences;
- inserting said polynucleotide into a vector in a microorganism;
- and
- expressing said peptide on the outer surface of said microorganism such that said single polynucleotide is capable of producing a plurality of different polypeptides.
Dependencies are inferred from the claim text.