Composite

Part:BBa_K364326:Experience

Designed by: Endre Károly Kristóf   Group: iGEM10_Debrecen-Hungary   (2010-10-20)


Ligand

Ecdysteroids

Ecdysteroids are insect moulting and sex hormones which include ecdysone and its homologues such as 20-hydroxyecdysone. Ecdysteroids also occur in other invertebrates where they can play a different role. They also appear in many plants mostly as a protection agents (toxins or antifeedants) against herbivore insects. These phytoecdysteroids have medicinal value and are part of herbal adaptogenic remedies like cordyceps.

Physiological functions of EcR

Pulses of 20-hydroxyecdysone occur during insect development, whereupon this hormone binds to the ecdysone receptor, a ligand-activated transcription factor found in the nuclei of insect cells. This in turn leads to the activation of many other genes, as evidenced by chromosomal puffing at over a hundred sites. Ultimately the activation cascade causes physiological changes that result in ecdysis (moulting).

Potential applications of GAL4-EcR LBD

By the addition of lipid ligands(ecdysteroids), the created system activates the expression of any kind of genes fused to UAS(upstream activation seqence)previously. Also can be used as a lipid sensor by activating reporter genes (e.g. Luciferase).

Applications of BBa_K364326

Picture of gel electrophoresis: Gal4-EcR in pCDNA3.1 resulting an insert of 1142 bp.
Picture of Western blot: Gal4-EcR composite construct is expressed after transferring it into pCDNA3.1
Picture of Western blot: Gal4-EcR composite construct is highly expressed after transferring it into pCDNA3.1
Test of pCDNA/pSB1A3-GAL4-PXR/EcR-LBD activity

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