Ribosome-mediated incorporation of peptides and peptidomimetics
AssigneeArizona Board of Regents on behalf of Arizona State University
InventorsSidney Hecht, Larisa (Liza) Dedkova, Rumit Maini, Sandipan Roy Chowdhury, Rakesh Paul
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Abstract
Modified ribosomes that were selected using a dipeptidyl-puromycin aminonucleoside are used to mediate site-specific incorporation of one or more peptides and peptidomimetics into protein in a cell free translation system. In addition, new fluorescent dipeptidomimetics have been synthesized and incorporated into proteins, as well as modified proteins containing one or more non-naturally occurring dipeptides.
Claims16
An Escherichia coli (E. coli) bacterial ribosome comprising a modified 23S ribosomal RNA sequence comprising modifications as compared to a wild-type counterpart, wherein the modifications consist of a first modified region (Region 1) and a second modified region (Region 2);
- (a) if the modification in Region 1 consists of UGCGUGG from position 2057-2063, the modification in Region 2 is selected from the group consisting of ACGAAG, CGCACG, CUAUGU, CGCAAU, or CUACAG from position 2502-2507;
- (b) if the modification in Region 1 consists of AGCGUGA from position 2057-2063, the modification in Region 2 is selected from the group consisting of CUGCGU, UGGCAG, AUCAGG, or AUCCGA from position 2502-2507;
- wherein the wild-type counterpart DNA sequence of the first unmodified region (Region 1) consists of the sequence GAAAGAC from position 2057 to 2063;
- wherein the wild-type counterpart DNA sequence of the second unmodified region (Region 2) consists of the sequence GAUGUC from position 2502-2507;
- and
- wherein the 23S ribosomal RNA is expressed from an engineered rrnB operon.
Dependencies are inferred from the claim text.