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Expanding the zebrafish genetic code through site-specific introduction of azido-lysine, bicyclononyne-lysine, and diazirine-lysine
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Syed, Junetha, Palani, Saravanan, Clarke, Scott T., Asad, Zainab, Bottrill, Andrew R., Jones, Alexandra M.E., Sampath, Karuna and Balasubramanian, Mohan K. (2019) Expanding the zebrafish genetic code through site-specific introduction of azido-lysine, bicyclononyne-lysine, and diazirine-lysine. International Journal of Molecular Sciences, 20 (10). 2577. doi:10.3390/ijms20102577
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Official URL: http://dx.doi.org/10.3390/ijms20102577
Abstract
Site-specific incorporation of un-natural amino acids (UNAA) is a powerful approach to engineer and understand protein function. Site-specific incorporation of UNAAs is achieved through repurposing the amber codon (UAG) as a sense codon for the UNAA, using a tRNACUA that base pairs with an UAG codon in the mRNA and an orthogonal amino-acyl tRNA synthetase (aaRS) that charges the tRNACUA with the UNAA. Here, we report an expansion of the zebrafish genetic code to incorporate the UNAAs, azido-lysine (AzK), bicyclononyne-lysine (BCNK), and diazirine-lysine (AbK) into green fluorescent protein (GFP) and glutathione-s-transferase (GST). We also present proteomic evidence for UNAA incorporation into GFP. Our work sets the stage for the use of AzK, BCNK, and AbK introduction into proteins as a means to investigate and engineer their function in zebrafish.
| Item Type: | Journal Article | |||||||||||||||
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| Divisions: | Faculty of Medicine > Warwick Medical School | |||||||||||||||
| Journal or Publication Title: | International Journal of Molecular Sciences | |||||||||||||||
| Publisher: | M D P I AG | |||||||||||||||
| ISSN: | 1422-0067 | |||||||||||||||
| Official Date: | 26 May 2019 | |||||||||||||||
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| Date of first compliant deposit: | 26 June 2019 | |||||||||||||||
| Volume: | 20 | |||||||||||||||
| Number: | 10 | |||||||||||||||
| Article Number: | 2577 | |||||||||||||||
| DOI: | 10.3390/ijms20102577 | |||||||||||||||
| Status: | Peer Reviewed | |||||||||||||||
| Publication Status: | Published | |||||||||||||||
| Access rights to Published version: | Open Access | |||||||||||||||
| RIOXX Funder/Project Grant: |
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