Characteristics of bacterial strains and plasmids utilized in this study
Strains/Plasmids | Characteristics | Sources |
---|---|---|
DH10B | F-, |
Invitrogen |
DH10B (Tn: |
DH10B containing chromosomal integration of |
Kim |
M15 (pREP4) | Nals, Strs, Rifs, Thi-, Lac-, Mtl-, F-, [pREP4 KmR], used for production of aaRS recombinant proteins | Qiagen |
Plasmids | ||
pACamG | CAT gene with an amber mutation at the 27th residue and |
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 |
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 |
This study |
pACamG-AzPheRS-1, -2, and, -3 | Vector expressing both |
This study |
pCNFRS | Vector expressing both AzPhe-incorporating aaRS derived from |
Miyake-Stoner |
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 |
Abbreviations: CAT, chloramphenicol acetyl transferase; aaRS, aminoacyl-tRNA synthetase;