Figure 3
fliC::lacZ expression monitors in vivo GreA reversal of transcription elongation arrest in ΔdksA ΔgreA host cells carrying pGreA derived plasmids. fliC-lacZ β-gal activities are made in the PRG18 strain (ΔdksA ΔgreA fliC::lacZ distal) bearing plasmids pTrc99a (pControl), pDNL278 (pGreA) or each of the 20 different pTrc-GreAMut. Cultures were grown in LB at 37 °C up to an OD600nm 1.5. The β-galactosidase activities shown are normalized to pGreA, set as 100% of antipause activity. Error bars represent SD from 2 biological replicates and 3 technical replicates. Mutant classes are colour coded as in Fig. 2c: Class I red, Class II green, Class III yellow.
