Team:ZJU-China/Hardware/MediumWave
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Medium Wave
For this year’s iGEM competition, our team has chosen to present Calcineurin-dependent response element (CDRE) (BBa_K2207021)and Mature serine protein from Paecilomyces lilacinus (BBa_K2207023) for the award of the basic part.
CDRE
CDRE，calcineurin-dependent response element, is a segment of DNA sequence which can be regulated by specific transcription factor. This TF is activated by calcineurin in the present of calcium ion. To create a promoter can be up-regulated by calcium influx，we replaced the upstream activating sequence(UAS) of CYC1 promoter with 4 CDREs. This promoter is designed for Saccharomyces cerevisiae and it depends on S.cerevisiae's endogenous calcineurin and calmodulin[1][2]. We choose mRFP as the report gene. We cultured the yeast in calcium-inducing medium and uninduced medium which contains a relatively low concentration of calcium ion (1xYPD medium).
Fig.1 (a) Relative fluorescent intensity of two groups. (b) Relative calcium content in two groups
Mature serine protein
Root-knot nematodes (Meloidogyne spp.), which are one of the most destructive nematodes, cause the loss of crop about 10%, serious as high as 75%. Like the insect cuticlehe, nematode eggshell consists mainly of proteins and chitins. The egg-parasitic fungus P.lilacinum secretes protease and chitinase to hydrolyze the nematode eggshell, so that the root knot nematodes cannot grow normally[3]. Among them, serine protases plays an important role in hydrolyzing the eggshell of root-knot nematodes.
Fig.2 Western-blot result
Table of the Basic Parts we submitted to the BioBrick Registry
| Name | Type | Description | Designer | Length(bp) | Submitted |
|---|---|---|---|---|---|
| BBa_K2207003 | Composite Parts | Trichoderma HR System I SOD-Tubulin dobule promoter | Yihe Zhang | 3155 | ✔ |
Table.OD600 reference point
Reference
[1] Cyert M S. Calcineurin signaling in Saccharomyces cerevisiae: how yeast go crazy in response to stress[J]. Biochemical and biophysical research communications, 2003, 311(4): 1143-1150.
