ActinoMation

ActinoMation facilitates a semi-automated, medium-throughput workflow for introducing recombinant DNA into Streptomyces spp. using the Opentrons robotics platform.

Key Features:

• Literate programming environment: ActinoMation uses Python in Jupyter Notebook as a protocol-creator for robotic workflows.
• Open-source protocol generation: The tool generates open-source protocols for the Opentrons OT-2 robotics platform.
• Conjugation efficiency validation: The workflow achieves conjugation efficiencies ranging from 1.21*10-5/0.0012% to 6.13*10-2/6.13% across tested strains and plasmids.

Scientific Applications:

• Streptomyces genetic engineering: ActinoMation enables medium-throughput robotic conjugation for introducing recombinant DNA into Streptomyces coelicolor (M1152 and M1146), S. albidoflavus (J1047), and S. venezuelae (DSM40230).
• Plasmid transformation: The workflow supports transformation with plasmids pSETGUS and pIJ12551.
• Metabolic engineering: The tool facilitates access to the metabolic potential of Streptomyces for antibiotic and bioactive compound production.

Methodology:

ActinoMation employs a literate programming approach using Python in Jupyter Notebook to create protocols for the Opentrons OT-2 robotics platform. The method was validated by transforming Streptomyces strains via conjugation and measuring conjugation efficiencies and false positive rates.