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Wet Lab
Overview
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Chassis
(http://2013.igem.org/Team:Cornell/project/wetlab/chassis)
Fungal Toolkit
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit)
Regulatory
Elements
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/regulatory_elements)
Selectable
Markers
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/selectable_markers)
Characterization
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/characterization)
Homologous
Constructs
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/homologous_constructs)
Biosafety
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/biosafety)
Protoplasting
(http://2013.igem.org/Team:Cornell/project/wetlab/protoplasting)
Carotenoids
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/carotenoids)
Limonene
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/limonene)
Antifungals
(http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit/antifungals)
BioBricks
(http://2013.igem.org/Team:Cornell/project/wetlab/biobricks)
Animation
(http://2013.igem.org/Team:Cornell/project/wetlab/animation)
Future Work
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Wetlab Overview
We are working with the fungus Ganoderma lucidum, a model organism for the class of wood-
rot fungi that is used in Ecovatives products. We have assembled a number of constructs for
the production of carotenoid pigmentation, fluorescent proteins, and antibiotic resistances that
we are seeking to transform by both random insertion and homologous recombination into the
organisms genome. These proof-of-concept tools are necessary to optimize our process so
that we can then obtain rigorous, reliable characterization data for the antifungal gene we are
seeking to introduce, a protein that specifically targets Aspergillus niger.
However, as Ganoderma does not have a well-standardized transformation protocol and takes
several days to grow to an appropriate density, we are also conducting much of our basic fungal
characterization work in Cochliobolus heterostrophus, a simpler fungus. In addition, we are
seeking to introduce a novel T7 viral regulation system into fungi. Similar systems have been
used successfully in mammalian cells, and this system would greatly expand the accessibility
of fungal genetic engineering beyond experienced mycologists. This system allows us to
conduct preliminary characterization within E. coli BL21-A1, a much simpler system for
acquiring quantitative data.
We are also implementing a number of biosafety measures within our toolkit, including a
recombination system that has previously been demonstrated to work in simpler fungi, which
will allow us to remove antibiotic resistance genes and targeted antifungal compounds before
the end of the production process, in order to prevent them from spreading into the
environment. The strain we are seeking to develop would be a huge boon to the economic
viability of Ecovatives sustainable biomaterials, as it would allow them to more easily maintain
the high levels of quality control that are necessary in scaled-up production.
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