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<h3><a target="_blank" href="https://2017.igem.org/Team:Bielefeld-CeBiTec/Project/toolbox/fusing">Fusing</a></h3> | <h3><a target="_blank" href="https://2017.igem.org/Team:Bielefeld-CeBiTec/Project/toolbox/fusing">Fusing</a></h3> | ||
<article> | <article> | ||
− | We explored the possibilities of a new method for highly specific protein terminus independent fusing based on the binding of 1,2‑aminothiols and the cyano group of cyanobenzothiazole (CBT). Therefore, we designed and synthetized a new amino acid by coupling the amino group of 6‑amino‑2‑cyanobenzothiazole and the carboxyl group of the side chain of aspartic acid. The result was N<sup>γ</sup>‑2‑cyanobenzothiazol‑6‑yl‑L‑asparagine (CBT‑Asp). As a counterpart, we used N<sup>ε</sup>‑L‑cysteinyl‑L‑lysine (CL), which contains a 1,2‑aminothiol group. This way, we created a pair of non‑canonical amino acids residues can bind covalently to each other under physiological conditions. We cloned the aaRS for CL (<a target="_blank" href="http://parts.igem.org/Part:BBa_K2201208">BBa_K2201208</a>) in pSB1C3 and used our modeling to predict 12 possible sequences of an aaRS able to incorporate the new amino acid CBT‑Asp (<a target="_blank" href="http://parts.igem.org/Part: | + | We explored the possibilities of a new method for highly specific protein terminus independent fusing based on the binding of 1,2‑aminothiols and the cyano group of cyanobenzothiazole (CBT). Therefore, we designed and synthetized a new amino acid by coupling the amino group of 6‑amino‑2‑cyanobenzothiazole and the carboxyl group of the side chain of aspartic acid. The result was N<sup>γ</sup>‑2‑cyanobenzothiazol‑6‑yl‑L‑asparagine (CBT‑Asp). As a counterpart, we used N<sup>ε</sup>‑L‑cysteinyl‑L‑lysine (CL), which contains a 1,2‑aminothiol group. This way, we created a pair of non‑canonical amino acids residues can bind covalently to each other under physiological conditions. We cloned the aaRS for CL (<a target="_blank" href="http://parts.igem.org/Part:BBa_K2201208">BBa_K2201208</a>) in pSB1C3 and used our modeling to predict 12 possible sequences of an aaRS able to incorporate the new amino acid CBT‑Asp (<a target="_blank" href="http://parts.igem.org/Part:BBa_K2201302">BBa_K2201302</a>). |
</article> | </article> | ||
</div> | </div> |
Revision as of 19:04, 1 November 2017
![](https://static.igem.org/mediawiki/2017/e/e9/T--Bielefeld-CeBiTec--title-img-toolbox.jpg)
Toolbox Overview
Short Summary
Analyzing
The structural analysis of a protein can be used to study distances between non-canonical amino acids (ncAAs) with Foerster Resonance Energy Transfer (FRET). This provides measuring distances between specific incorporated amino acids in the target protein. To incorporate the ncAAs we provide three different tRNA/aminoacyl-synthetases (BBa_K2201201, BBa_K2201202, BBa_K2201203) which incorporate in response to the amber or the less used leucine codon. To demonstrate this tool we also developed a prion detection assay.