sils presentation
DESCRIPTION
TRANSCRIPT
Genetically Engineered Mycelium for Biomaterials
DevelopmentBy Arun Chakravorty and Eric Holmes
• International Genetically Engineered Machines
• Standardization of genetic parts using “BioBricks”
• Development and expansion of genetic parts registry
• Annual competitions in October and November
• Developed collaboration with Ecovative.
• Company that creates Styrofoam substitute using fungal mycelium
• Hope to use synthetic biology to improve marketability
Chassis
• Ganoderma lucidum
• Higher order basidiomycete
• Sequenced in 2012• Reported to have
medicinal properties
Goals• Improve product via introduction of
several biological plasmid constructs.• Antifungal constructs to eliminate fungal contaminants
o Provide fungal species opportunity to outcompete fungal contaminants
• Carotenoid pigment pathways to make product more appealing
• Develop fungal toolkit
Homologous Recombination• Integrate linearized plasmids via natural
homologous recombination
Figure from Gene Bridges
Gibson Assembly• Used for cloning in genes with internal cut sites• Uses polymerase, 5’ exonuclease, and ligase in simultaneous reaction
Photo from Integrated DNA Technologies
Constructs
Site-Directed Mutagenesis• Introduce silent mutations• Used on genes with internal restriction sites
Image from Agilent Technologies
Thermal Cycling
DpnI Digestion
Transformation
Promoter Problems• Fungal promoters are very long
o E.coli has a check system where it splices out potential duplications
• Very few identified Ganoderma promoters• Potential solution: use T7 bacteriophage
promoter/polymerase
Image credit: Thomas Splettstoesser
Constructs
Acknowledgments
• Weill Institute• Cornell Institute for
Biotechnology & Life Sciences Technology
• College of Engineering• Turgeon Lab• Corning• Geneious• The entire Cornell iGEM
team
Thank you for listening!