Korean Journal of Microbiology

Table. 1.

Characteristics of bacterial strains and plasmids utilized in this study

Strains/Plasmids Characteristics Sources
Escherichia coli strains
DH10B F-, mcrA, Δ (mrr-hsdRMS-mcrBC), Φ80d lacZΔM15, ΔlacX74, deoR, recA1, endA1, araD139, Δ (ara, leu)7697, galU, galK, λ-rpsL, nupG Invitrogen
DH10B (Tn:lacZam) DH10B containing chromosomal integration of lacZam-KmR cassette Kim et al. (2009)
M15 (pREP4) Nals, Strs, Rifs, Thi-, Lac-, Mtl-, F-, [pREP4 KmR], used for production of aaRS recombinant proteins Qiagen

pACamG CAT gene with an amber mutation at the 27th residue and E. coli mutant initiator tRNA2fMet (trnfMT2:A71/T35A36/G72) called fMamtRNACUA, pACYC184 frame, TetR Chow and RajBhabdary (1993)
pQE-30 Commercial bacterial expression vector, AmpR Qiagen
pQE-YRS Sc YRS expression vector. CmR gene (CAT) was removed from pQE-30 and Sc YRS gene was cloned with N-terminal 6xHis tag Tekalign et al. (2018)
pQE-AzPheRS-1, -2, and -3 Vector expressing AzPheRS proteins that were generated by site-directed mutagenesis with pQE-YRS as template This study
pACamG-YRS Vector expressing both Sc YRS and fMamtRNACUA. Sc YRS gene was subclone into pACamG This study
pACamG-AzPheRS-1, -2, and, -3 Vector expressing both Sc AzPheRS mutants and fMamtRNACUA. Sc AzPheRS genes were subcloned into pACamG This study
pCNFRS Vector expressing both AzPhe-incorporating aaRS derived from Mj YRS and suppressor tRNA, araBAD promoter, CmR. Miyake-Stoner et al. (2010)
pBad-sfGFP-150TAG Reporter plasmid, superfold GFP gene with an amber mutation at the 150th residue, araBAD promoter, 6xHis tag at the C-terminus, AmpR Miyake-Stoner et al. (2010)

Abbreviations: CAT, chloramphenicol acetyl transferase; aaRS, aminoacyl-tRNA synthetase; Sc, Saccharomyces cerevisiae; Mj, Methanocaldococcus jannaschii; CmR, chloramphenicol resistance; AmpR, ampicillin resistance; TetR, tetracycline resistance.

Korean J. Microbiol. 2024;60:1-11 https://doi.org/10.7845/kjm.2024.4009
© 2024 Korean J. Microbiol.