From Zhang et al. (2015)
Bioengineered Acaricide from African Biodiversity Moola Mutondo Designer Cells Node
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Abstract African plants contain many cyclotides, ultra‑stable peptides with major potential as drugs and biopesticides, but functional screening of this diversity is limited. The goal is to build a programmable biosensor platform to screen African plant germplasm for cyclotides that modulate a defined bacterial regulatory pathway. The project tests the hypothesis that cyclotides or related peptides which interact with LacI will cause measurable changes in a LacI‑controlled GFP reporter circuit. Specific aims are: (1) design and assemble a plasmid encoding a constitutive LacI cassette and a LacI‑regulated sfGFP reporter, (2) establish a cell‑free or E. coli assay compatible with lyophilised plant extracts, and (3) implement a small‑scale, automated screen (e.g., on an Opentrons robot) to identify extracts that reproducibly alter GFP output. Methods include DNA design and synthesis, cell‑free expression or bacterial culture, plant extract preparation, automated liquid handling, and plate‑reader fluorescence measurements.