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. 2010 Nov 9;39(3):e12. doi: 10.1093/nar/gkq1070

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — Isolating AHL Inverters and NAND-gates from a single rbs library. (A) Library design. The six bases (N₆ in figure) corresponding to rbs of luxR gene were randomized. (B) The phenotype variation of the circuit library before (left), after the ‘AHL-inverter selection’ (middle), and after the ‘NAND-gate selection’ (right). (C) Distribution of the rbs scores in the variants from original (black filled diamonds), AHL inverter-selected (red filled circles), and NAND gate-selected (blue filled circles) libraries. X-axis represents the free binding energy of these rbs sequences (ΔG_binding) and anti-rbs sequence, calculated using the RNA duplex software component of the Vienna RNA Package (45). Rbs scores for Y-axis were calculated using RBS Calculator (14). The sequence AGGAGG (perfect match to the anti-rbs) was independently constructed by site-directed mutagenesis. It was found to behave as an AHL inverter.