| 초록 |
In this study, a new aminoacyl-tRNA synthetase (AARS) was developed for the expression of recombinant protein with non-natural amino acid. For this, 4-Azido-L-phenylalanine (AzF) was used as a non-natural amino acid, and a mutant library for AARS, derived from Metanosaeta concilii, was prepared through error prone PCR. Each AARS mutant gene was inserted into the pEVOL-pAzF vector. In addition, the sfGFP (superfolded green fluorescence protein) gene in which codon 140 was substituted with amber was inserted into the pSEVA631PT vector as a reporter gene, and both vectors were transformed into Escherichia coli DH10B. Fluorescence-activated cell sorting (FACS) was used to screen strains that fluoresce only when AzF was present in the medium. The selected mutant strain exhibited fluorescence of 33-60% of the control AARS which was derived from Methanococcus jannaschii. |
