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Figure 3 | Journal of Biological Engineering

Figure 3

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

Figure 3

Excision and selection for BioScaffold removal. Here, the prototype BioScaffold is present in a test circuit, with sequence "promoter-scar-BioScaffold part-scar-GFP." It is cut with the enzyme PsrI and then replaced with a RBS sequence. The BioScaffold test part contains a RFP expression circuit surrounded by two PsrI recognition sites; thus, cells that contain the BioScaffold part, such as the cells that contain the test circuit, exhibit red fluorescence. If PsrI excises the BioScaffold part from the test circuit and an RBS is ligated across the open gap, then the sequence "promoter-scar-RBS-GFP" is obtained. Cells that contain this sequence exhibit green flourescence, where the strength of the fluorescence depends on the RBS used. To show the flexibility of the BioScaffold, it was also replaced with a "RBS-MBP-GS" sequence to create the sequence "promoter-scar-RBS-MBP-GS."

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Journal of Biological Engineering

ISSN: 1754-1611