Difference between revisions of "Part:BBa K2201205"

Latest revision as of 16:07, 1 November 2017

Cysteinyl-lysinyl-tRNA Synthetase

This cysteinyl-lysinyl-tRNA synthetase is based on the work of Nguyen et al. (2011). It is originated from the pyrrolysyl-tRNA synthetase of Methanosarcina barkeri containing a C313V mutation.
N^ε-L-cysteinyl-L-lysine (CL) is an amino acid containing a free 1,2-aminothiol group at its side chain. 1,2-aminothiols are an important part of the synthesis of D-Luciferin. D-Luciferin is the substrate of the luciferase of the firefly Photinus pyralis. The synthesis of D-Luciferin is based on a condensation reaction between CL and a cyanobenzothiazole derivative resulting in a covalent bond between two thiazole residues. We designed and synthesized the novel amino acid N^γ‑2‑cyanobenzothiazol‑6‑yl‑L‑asparagine which contains 6-amino-2-cyanobenzothiazole at its side chain. Additionally, we showed that both amino acids are able to undergo the mentioned condensation reaction enabling a system for highly specific binding between peptides and enzymes.
So this synthetase is an important part of our fusing tool. Together with the CLtRNA (K2201206) we developed a aminoacyl-tRNA synthetase/tRNA pair (K2201208) to incorporate CL.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 492
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2201205 short</partinfo>
-This cysteinyl-lysinyl-tRNA Synthetase is based on the work of Nguyen <i>et al</i>. (2011). It is originated from the pyrrolysyl-tRNA synthetase of <i>Methanosarcina barkeri</i> containing a C313V mutation&#1102;
+This cysteinyl-lysinyl-tRNA synthetase is based on the work of Nguyen <i>et&nbsp;al</i>. (2011). It is originated from the pyrrolysyl-tRNA synthetase of <i>Methanosarcina barkeri</i> containing a C313V mutation.
+<br>
-N<sup>&#949;</sup>-L-cysteinyl-L-lysine is an amino acid containing a free 1,2-aminothiol group at its side chain. 1,2-aminothiols are an important part of the synthesis of D-Luciferin. D-Luciferin is the substrate of
+N<sup>ε</sup>-L-cysteinyl-L-lysine (CL) is an amino acid containing a free 1,2-aminothiol group at its side chain. 1,2-aminothiols are an important part of the synthesis of D-Luciferin. D-Luciferin is the substrate of
 the luciferase of the firefly <i>Photinus pyralis</i>. The synthesis of D-Luciferin is based on a condensation reaction between CL and a cyanobenzothiazole derivative resulting in a covalent bond between two thiazole
-residues. We designed and synthesized the novel amino acid <i>N</i><sup>&#947;</sup>&#x2011;2&#x2011;cyanobenzothiazol&#x2011;6&#x2011;yl&#x2011;L&#x2011;asparagine which contains 6-amino-2-cyanobenzothiazole at its side chain.
+residues. We designed and synthesized the novel amino acid <i>N</i><sup>γ</sup>&#x2011;2&#x2011;cyanobenzothiazol&#x2011;6&#x2011;yl&#x2011;L&#x2011;asparagine which contains 6-amino-2-cyanobenzothiazole at its side chain.
 Additionally, we showed that both amino acids are able to undergo the mentioned condensation reaction enabling a system for highly specific binding between peptides and enzymes.
+<br>
 So this synthetase is an important part of our <html><a href="http://2017.igem.org/Team:Bielefeld-CeBiTec/Project/toolbox/fusing">fusing tool</a></html>. Together with the CLtRNA
 (<html><a href="https://parts.igem.org/Part:BBa_K2201206">K2201206</a></html>) we developed a aminoacyl-tRNA synthetase/tRNA pair (<html><a href="https://parts.igem.org/Part:BBa_K2201208">K2201208</a></html>) to incorporate