Evolution of Context Dependent Regulation by Expansion of Feast/Famine Regulatory Proteins [expression]

We validated context dependent regulation by two FFRPs AsnC and VNG1237C by deleting the FFRP and testing the effect on downstream target genes. For validtion of AsnC condition specific regulation we grew wild-type and delta-AsnC in the presence of 4mM paraquat (PQ) and samples at 1 and 160 minutes post addition of this sub-lethal dose of PQ. For validation of VNG1237C we grew wild-type and delta-VNG1237C in a standard growth curve sampling at OD 0.18 and 1.15. The asnC deletion strain (∆asnC) and the control ∆ura3 strain were grown in 4mM paraquat (PQ) and were sampled at 1 and 160 minutes after PQ addition. Sampling timing for ∆VNG1237C was designed such that the starting ODs were as similar as possible and once an OD of 1.17 was reached for ∆ura3 the ∆VNG1237C was sampled at this same time point.