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This avoids the handling of possibly toxic, free cellular metal ions and provides a coordinated involvement in metabolic pathways such as specific incorporation in metalloproteins or detoxification.
Previously, we have developed genetically encoded unnatural amino acid (3,5-difluorotyrosine, F2Y) to implement site specific incorporation in ETK-CD (Li et al., 2013).
Specific incorporation of the c.1114C>T mutation was confirmed by sequencing insertion-specific PCR products.
As evident from figure 4B, again specific incorporation of radioactivity into different peptides was observed.
Note that the antisense sequence has mismatches with the miR-10 sense strand at its 3' end resulting in the specific incorporation of the sense miR-10 strand in the microRNA silencing complex.
Extracts from these cells were incubated with a 34-bp duplex substrate containing a single uracil residue at position 16 in a reaction containing [32P]dCTP and chain terminator ddGTP (Fig. 9A) to measure pol β specific incorporation.
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The site-specific incorporation (SSI) of UAAs utilizes nonsense codons to incorporate one or a few UAAs into a single or multiple defined locations in a protein.
Techniques for position-specific incorporation of non-natural amino acids in an in vitro protein synthesizing system are described.
The method is versatile and allows stoichiometric and active site-specific incorporation of any one of many molecular probes.
This review briefly recaps the residue- and site-specific incorporation techniques for non-canonical amino acids in yeast.
Residue-specific incorporation of non-canonical amino acids into proteins allows facile alteration and enhancement of protein properties.
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