Team:CIEI-China/Notebook
We came up with the idea about using salt tolerant yeast to dispose kitchen waste which have high osmotic pressure. Then we started to search the papers related to our research.
We found three genes that are possible for our goal: SpTPS1, ScTPS1, and gltB.
And then we began to prepare the materials we need in the following experiments.
We finished the design of nine biobricks with different functions.
Designing and editing of the experiment plan.
Transformation of PUC57 plasmid into DH5α.
Culture the E.coli.
Preparation of plasmid DNA by Alkaline Lysis with SDS;
Digestion (XbaⅠ, SpeⅠ).
Recycle the electrophoresis product.
Linearization of the standard carrier.
Ligate the genes into the standard carrier.
Transform the standard carrier into DH5α E.coli;
Culture the E.coli and prepare the plasmid.
Design the primers of the three genes: SpTPS1, ScTPS1, and gltB.
PCR PrimeStar DNA polymerase.
Recycle the plasmids from PCR products.
Digest the PCR products.
Electrophoresis and recycling.
Ligation;
We successfully ligated the three target genes in to the vector pPIC9K to get the devices.
Transform the vector into DH5α E.coli.
Culture the E.coli.
Extract the devices.
Electrophoresis of six completed devices: intracellular and extracellular expression of the three target genes. Recycle the devices from the gels.
Transform the devices into GS115 competent yeast cells.
Sequencing the genes we transformed into the yeast.
Induce experiments by using methanol.
SDS-Page.
Test the survive rate of the yeast in different salinity.
Test the decomposition rate of the waste.
