Naslov
Optimization of unnatural amino acid incorporation via amber suppression in Caenorhabditis elegans

Autori
Ljubišić, Tea

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, diplomski rad, diplomski

Fakultet
Prirodoslovno-matematički fakultet

Mjesto
Zagreb

Datum
25.02

Godina
2015

Stranica
57 + xvi

Mentor
Chin, Jason W. ; Rokov Plavec, Jasmina

Neposredni voditelj
Radman, Inja

Ključne riječi
amber suppression/ Caenorhabditis elegans/ E55D release factor 1/ orthogonal aminoacyl-tRNA synthetase/tRNA system/ photocaged tyrosine/ pyrrolysyl-tRNA synthetase/ pyrrolysyl-tRNA/ U25C tRNACUA

Sažetak
Genetic code expansion is a group of technologies which aim to introduce non-natural functionalities into the proteome in a genetically directed, site-specific fashion. One of the methods to achieve this is through amber suppression with an orthogonal aminoacyl-tRNA synthetase/tRNA pair, such as the evolved pyrrolysyl-tRNA synthetase/pyrrolysyl-tRNA from methanogenic archaea, which specifically recognizes only the desired unnatural amino acid. Many designer amino acids have been incorporated by this method, e.g. photocaged amino acids. This thesis addressed the incorporation of a novel unnatural amino acid, photocaged tyrosine (PCY), in model organism Caenorhabditis elegans, as well as strategies for the improvement of C. elegans amber suppression for previously incorporated photocaged lysine (PCK). In order to increase the amber suppression yields, we attempted to engineer the competing eRF1 and the pyrrolysyl- tRNA itself, which were reported recently in the literature as optimization targets in other systems. We didn’t observe the incorporation of PCY and no significant change in suppression levels were detected for the use of pyrrolysyl- tRNA or eRF1 tested variants.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija

POVEZANOST RADA