Difference between revisions of "Team:TU Dresden/Improve"

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<p>The SpyCatcher is a highly functional Tag intended for translational fusion to a target protein. Unfortunately its size (116 amino acid) is quite large and can therefor hinder the native folding and successfull secretion of the protein of interest. We submitted a minimized version of the <a target="_blank"href="http://parts.igem.org/Part:BBa_K2273015">SpyCatcher</a> to the registry to ensure the effective secretion necessary for our project. The mini. SpyCatcher is 32 amino acids shorter but still fully functional, which was proven by our team (have a look at our <a href="https://2017.igem.org/Team:TU_Dresden/Project/Secretion">results</a>) as well as by the <a target="_blank"href="https://www.ncbi.nlm.nih.gov/pubmed/24161952">researchers</a> that proposed the minimization.</p>
 
<p>The SpyCatcher is a highly functional Tag intended for translational fusion to a target protein. Unfortunately its size (116 amino acid) is quite large and can therefor hinder the native folding and successfull secretion of the protein of interest. We submitted a minimized version of the <a target="_blank"href="http://parts.igem.org/Part:BBa_K2273015">SpyCatcher</a> to the registry to ensure the effective secretion necessary for our project. The mini. SpyCatcher is 32 amino acids shorter but still fully functional, which was proven by our team (have a look at our <a href="https://2017.igem.org/Team:TU_Dresden/Project/Secretion">results</a>) as well as by the <a target="_blank"href="https://www.ncbi.nlm.nih.gov/pubmed/24161952">researchers</a> that proposed the minimization.</p>
 
<h3>Codon adaption</h3>
 
<h3>Codon adaption</h3>
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<p>The original part was used in <i>E. coli</i> but the mini. SpyCatcher was implemented in <i>B. subtilis</i> so the code was adapted to its codon usage. The sequence was proposed by [https://www.ncbi.nlm.nih.gov/pubmed/28230977 researchers]that investigated the secretion of SpyCatcher fusion proteins with <i>B. subtilis</i>.  </p>
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<p>The original <a target="_blank"href="http://parts.igem.org/Part:BBa_K1159200">part</a> was used in <i>E. coli</i> but the mini. SpyCatcher was implemented in <i>B. subtilis</i> so the code was adapted to its codon usage. The sequence was proposed by <a target="_blank"href="[https://www.ncbi.nlm.nih.gov/pubmed/28230977">researchers</a> that investigated the secretion of SpyCatcher fusion proteins with <i>B. subtilis</i>.  </p>
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<p><b>His-Tag</b>:</p>
+
<h3>His-Tag</h3>
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The mini. SpyCatcher was also submitted to the [http://parts.igem.org/Part:BBa_K2273171 registry] with an added C-terminal His-Tag to purify the protein constructs.
+
<p>The mini. SpyCatcher was also submitted to the <a target="_blank"href="http://parts.igem.org/Part:BBa_K2273171">registry</a> with an added C-terminal His-Tag to purify the protein constructs.</p>
  
 
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Revision as of 17:07, 30 October 2017

The minimized SpyCatcher

Minimization

The SpyCatcher is a highly functional Tag intended for translational fusion to a target protein. Unfortunately its size (116 amino acid) is quite large and can therefor hinder the native folding and successfull secretion of the protein of interest. We submitted a minimized version of the SpyCatcher to the registry to ensure the effective secretion necessary for our project. The mini. SpyCatcher is 32 amino acids shorter but still fully functional, which was proven by our team (have a look at our results) as well as by the researchers that proposed the minimization.

Codon adaption

The original part was used in E. coli but the mini. SpyCatcher was implemented in B. subtilis so the code was adapted to its codon usage. The sequence was proposed by researchers that investigated the secretion of SpyCatcher fusion proteins with B. subtilis.

His-Tag

The mini. SpyCatcher was also submitted to the registry with an added C-terminal His-Tag to purify the protein constructs.