Team:Hannover/Parts

From 2014.igem.org

Revision as of 08:02, 1 October 2014 by Anke (Talk | contribs)

<groupparts>iGEM013 Hannover</groupparts>

Concept

Our goal was to engineer a transgenic plant, which is able to bind heavy metals like cadmium, arsenic, zinc and copper. Therefor metal binding proteins were fused (called top four metal binding protein, T4MBP). To avoid cytotoxic effects on the plant caused by the heavy metals we decided to place T4MBP to the cell exterior. With a secretion signal (Expa4) the T4MBP is directed to the extracellular space. The protein is tethered to the cell surface by using a cellulose binding domain (CBD). Plant cells have a cell wall which consists of cellulose. Via the cellulose binding domain the protein is attached to the cell wall.

Expa4 secretion signal, localizes to extracellular space (BBa_K1478000)

Coding region for first 20 aminoacids of plant protein Exansin4. Secretion signal for plants, when fused to a protein directs the protein to the extracellular space. Derived from the signal sequence of Expansin4 protein from Arabidopsis thaliana (NP_181500). The sequence was optimized for Escherichia coli and Arabidopsis thaliana codon usage. Analyzes showed that the part works.

1. Cutler, S. R.; Ehrhardt, D. W.; Griffitts, J. S.; Somerville, C. R. (2000): Random GFP::cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency. In Proceedings of the National Academy of Sciences 97 (7), pp. 3718–3723. DOI: 10.1073/pnas.97.7.3718.