Fig. 2: Expanding tunability of the cBUFFER through RBS strength and CsrB-CsrA affinity modulation. | Nature Communications

Fig. 2: Expanding tunability of the cBUFFER through RBS strength and CsrB-CsrA affinity modulation.

From: Rewiring native post-transcriptional global regulators to achieve designer, multi-layered genetic circuits

Fig. 2

A Workflow for tuning cBUFFER expression. To tune CsrA-CsrB affinity, mutant CsrB sequences containing mutations to known CsrA binding sites were tested. To vary RBS strength, all 1024 five nucleotide space sequences were computationally screened, resulting in 4 additional optimized sequences. B Time course of cBUFFER systems with variable CsrB sRNAs; WT CsrB (Blue), High-affinity CsrB (Dark Blue), and Low-affinity mutant CsrB L2 (Light Blue). C Time course of cBUFFER systems with variable RBS strengths derived from the computational screen. D Relative fluorescence for all combinations of RBS-CsrB cBUFFER systems, two hours post-induction. Samples were grown in biological triplicate, and data presented are the mean values +/- the standard deviation, represented as the error bars.

Back to article page