Team:CCU Taiwan/Improve

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Improved Parts

Our improved part is mainly from the TU Munich iGEM Team in 2012. We used Thaumatin for the experiment. Thaumatin is a single chain with 207 amino acids residues and eight disulfide bonds and a molecular weight of 22.2 kDa. It is highly water soluble, stable at heating (not for cooking, bakery, etc.) and stable under acidic conditions. The production of thaumatin is induced by an attack upon the plant by viroid pathogens.

We will transform BBa_K801080 to smd1168. Hope to increase the amount of gene expression

Type: characterization/ measurement

Part: BBa_K801080--Prepro-Thaumatin, yeast codon optimized

Part source: 2018 Kit Plate 6 Well: 16 I

Description:

Thaumatin is a single chain with 207 amino acids residues and eight disulfide bonds and a molecular weight of 22.2 kDa. It is highly water soluble, stable at heating (not for cooking, bakery, etc.) and stable under acidic conditions. The production of thaumatin is induced by an attack upon the plant by viroid pathogens.

Experiment:

Express Thaumatin using pGAPZ A transformation in Pichia strain SMD1168, compara the expression level of Thaumatin.

1. Digest pGAPZ A and BBa_K801080 with NotI.
2. Ligation
3. Transformation into E.coli
4. Kit-free plasmid miniprep
5. Transformation into SMD1168
6. Analysis by Quantitative Western Blots