*Seminar Series: IMB Chemistry and Structural Biology Division Seminar
*Location: Seminar Room 3.142, QBP Bldg 80, UQ St Lucia
*Date: 14/09/2017
*Time: 12:30

Speaker #1 details
*Title of talk: Incorporation of unnatural amino acids into peptides and proteins via sense codon reassignment
*Speaker's name: Zhenling Cui
*Speaker's organisation: IMB
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Talk Abstract: Incorporation of unnatural amino acids (uAAs) via codon reassignment is a powerful approach for introducing novel chemical and biological properties to recombinant polypeptides. However, the site-selective incorporation of multiple uAAs into polypeptides is hampered by the limited number of reassignable nonsense codons. To address this limitation we set out to orthogonalize several of the sense codons. We established a quantitative in vitro peptide translation assay that allows us to test the ability of synthetic tRNAs to decode any of 61 sense codons. The majority of synthetic Escherichia coli tRNAs could support protein translation. By formulating a semisynthetic tRNA complement or by using DNA-hybridization chromatography, we obtained an E.coli in vitro translation system devoid of different tRNA isoacceptors. Such system is dependent on the addition of synthetic tRNAs for the chosen sense codon to restore its translational activity. This allows multisite-selective uAAs incorporation when combined with amber-suppression. We successfully applied this approach to production of the calmodulin protein with FRET-forming fluorescent probes. The presented approach is universal and can be expanded to other sense codons and cell-free systems.
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Speaker #2 details
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Other Details:
*Host name: Kirill Alexandrov
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*Host email: k.alexandrov@imb.uq.edu.au