CHEM 451 Lecture Notes - Lecture 7: Aminoacyl-Trna, Phosphoserine, Transfer Rna
Document Summary
Slide 1 - incorporation of unnatural amino acids. Engineering paper, want to use unnatural amino acids. Compare insertion by ribosome with natural amino acids vs insertion of unatural a. a (orthogonal pair and amino acyl trna synthetase) > orthogonal = doesn"t interfere with other mechanisms, trna doesn"t get charged by endogenous synthetases. Overall goal of paper = integrate phosphoserine in a specific spot, not done in e. coli. E. coli has lots of phosphorylation sites, but this modification is post translational of mammalian proteins. Modifications, when you make protein e. coli, it"s hard to engineer system to put them in. No synthetase to charge with cysteine, so it can use phosphoserine then transfer it with s and another enzyme. Key to this paper to know what combo of synthetase/ enzymes replace trna synthetase for an amino acid on methane producing bacteria, because steps is to engineer synthetase - this is non existent in e. coli.
