Fig. 6.
(a) The BCAM0271-2 gene pair constitutes a type II TA system. Overnight cultures of E. coli strains expressing BCAM0272 or BCAM0271-2 were serially diluted (10−3 to 10−8, left to right). Dilutions were spotted on LBA with repressor (glucose 0.2%) and inducer (arabinose 0.2% (w/v) or rhamnose 0.2% (w/v)). (b). Synthesis of GFP-StrepII reporter protein expressed from the T7 promoter in an in vitro transcription-translation system in presence of BCAM0272 with (+) and without (−) acetyl-Coenzyme A. Products of reaction resolved by SDS-PAGE and visualised by Western Blot with anti-strepII-tag antibodies. (c) Acetylation of tRNA pool in an in vitro transcription-translation system by different GNAT toxins with (+) and without (−) [14C]acetyl-Coenzyme A. RNAs resolved by native PAGE and stained with methylene blue (top panel), gel was then dried and exposed to phosphor storage screen (bottom panel).