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Figure 4 | Journal of Biological Engineering

Figure 4

From: Introduction of customized inserts for streamlined assembly and optimization of BioBrick synthetic genetic circuits

Figure 4

GFP expression levels created by the different RBSs. Sequence verified clones of the form "promoter-scar-RBS-GFP" (a) were analyzed visually (b) and using flow cytometry (c). The inserted RBSs were expected to drive relative translation initiation rates of 10, 100, 1,000, 10,000, and 100,000 for the downstream sequence GFP with the cell fluorescence proportional to the translation initiation rate (a). Qualitative visual assessment revealed green fluorescent intensity commensurate with the expected values (b), although colony thickness can influence perceived intensity. Simultaneously transformed colonies of RBS10, RBS100, and RBS1,000 appeared light green, while RBS10,000 appeared green and RBS100,000 appeared bright green (b). Quantitative assessment using flow cytometry data revealed GFP intensity levels commensurate with expected values, except for the higher than expected translation initiation of GFP driven by RBS10 (c).

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