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

 
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<h1 class="box-heading"> The minimized SpyCatcher</h1>
 
<h1 class="box-heading"> The minimized SpyCatcher</h1>
 
<h3>Minimization</h3>
 
<h3>Minimization</h3>
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<p>The SpyCatcher is a highly functional Tag, that can interact with its protein partner: the SpyTag. Upon interaction, additional C- or N-terminally translational fused proteins are brought in close proximity and can be linked together via this Tag system. Unfortunately the size of the SypCather (116 amino acid) is quite large and can therefore hinder the native folding of a linked 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>
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<p>The SpyCatcher is a highly functional Tag, that can interact with its protein partner: the SpyTag. Upon interaction, additional C- or N-terminally translational fused proteins are brought in close proximity and can be linked together via this Tag system. Unfortunately the size of the SpyCatcher (116 amino acid) is quite large and can therefore hinder the native folding of a linked protein of interest. We submitted a minimized version of the Spy Catcher (<a target="_blank"href="http://parts.igem.org/Part:BBa_K2273015">mini. 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>
 
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<h3>Codon adaption</h3>
 
<h3>Codon adaption</h3>
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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 thought to be implemented throughout our project in <i>B. subtilis</I>. Therefore we codon adapted the sequence for this purposes. The sequence was proposed by <a target="_blank"href="https://www.ncbi.nlm.nih.gov/pubmed/28230977">researchers</a> that investigated the secretion of SpyTag/SpyCatcher system in <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>Escherichia coli</i> but the mini. SpyCatcher was thought to be implemented throughout our project in <i>Bacillus subtilis</I>. Therefore we codon adapted the sequence for this purposes. The sequence was proposed by <a target="_blank"href="https://www.ncbi.nlm.nih.gov/pubmed/28230977">researchers</a> that investigated the secretion of SpyTag/SpyCatcher system in <i>B. subtilis</i>.  </p>
 
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<h3>His-Tag</h3>
 
<h3>His-Tag</h3>
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<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 protein constructs when SpyTag/SpyCatcher link together.</p>
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<p>The mini. SpyCatcher was also submitted to the <a target="_blank"href="http://parts.igem.org/Part:BBa_K2273017">registry</a> with an added C-terminal His-Tag to purify protein constructs when SpyTag/SpyCatcher link together.</p>
  
 
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Latest revision as of 01:26, 2 November 2017

Our best improved part

The minimized SpyCatcher

Minimization

The SpyCatcher is a highly functional Tag, that can interact with its protein partner: the SpyTag. Upon interaction, additional C- or N-terminally translational fused proteins are brought in close proximity and can be linked together via this Tag system. Unfortunately the size of the SpyCatcher (116 amino acid) is quite large and can therefore hinder the native folding of a linked protein of interest. We submitted a minimized version of the Spy Catcher (mini. 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 Escherichia coli but the mini. SpyCatcher was thought to be implemented throughout our project in Bacillus subtilis. Therefore we codon adapted the sequence for this purposes. The sequence was proposed by researchers that investigated the secretion of SpyTag/SpyCatcher system in B. subtilis.

His-Tag

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