Fig. 2: IBM as a universal method to exhaust split sites for AND and NAND logic gate engineering. | Nature Communications

Fig. 2: IBM as a universal method to exhaust split sites for AND and NAND logic gate engineering.

From: A systematic approach to inserting split inteins for Boolean logic gate engineering and basal activity reduction

Fig. 2

a Any transcription factor with a function that can be wired to an assay-friendly output could be subjected to IBM for logic gate engineering. b–d Intein-bisection maps for TetR (b, 3 seams identified), SrpR (c, 4 seams), and ECF20 (d, 3 seams). Split clones of TetR and SrpR (or ECF) achieved major off (or on) activities when both the N-lobes and the C-lobes were present. y locations and error bars are mean and SD of median fluorescence from experiments performed on three different days (n ≥ 3, see Supplementary Data 2, sheet “sample_sizes” for exact values of n). Vertical dashed lines bound the permitted transposition window and horizontal dashed lines bound the ranges of activities that could be attained by the split proteins. DMF dimethylformamide, DAPG 2,4-Diacetylphloroglucinol. d By-products of IBM revealed a truncatable region of ECF20 which when removed did not adversely affect activation. b–d See Supplementary Fig. 10–12 for site distributions and activities at 5 h (TetR and ECF20) and 24 h (SrpR). See Supplementary Fig. 9 for explanations of controls (leftmost subplots).

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