An easy tool to monitor the elemental steps of in vitro translation via gel electrophoresis of fluorescently labeled small peptides

Antibiotics' mechanism of action might be addressed by BODIPY-peptides separation by urea PAGE. Shown are the products of MF₂-coding mRNA in vitro translation in the presence of BPY-M-tRNA^fMet and antibiotics indicated above the lanes. (A) No post-treatment of the translation mixtures. (B) Translation mixtures after synthesis were treated with 1 M NaHCO₃. Lane 1 corresponds to the BPY-M-tRNA^fMet. DMSO 1% (2) corresponds to a positive no antibiotic control, (Ths) thiostrepton (3), (Ery) erythromycin (4), (Tyl) tylosin (5), (Pur) puromycin (6), (Madu) madumycin (7), (Eta) etamycin (8), (Ret) retapamulin (9), (BlaS) blasticidin S (10). BPY is marked with a green oval—BODIPY label; black circle—methionine; gray circle—phenylalanine residue; blue oval—tRNA^fMet or tRNA^Phe.