Difference between revisions of "Part:BBa K3187028"
Line 3: Line 3:
 
<partinfo>BBa_K3187028 short</partinfo>
 
<partinfo>BBa_K3187028 short</partinfo>
  
−
<p>Sortase A7M (SrtA7M) is a calcium-independent transpeptidase derived from the enzyme Sortase A from Staphylococcus aureus.1 Similar to Sortase A, SrtA7M connects two peptides covalently, one with a poly-glycine (poly-G) at the N-terminus and one with LPXTG motif at the C-terminus. It acts by cleaving between Gly and Thr of the LPXTG motif and catalysing the formation of an amide bond to the poly-G sequence.2 For the expression and purification of SrtA7M, a part containing T7 promoter, a lac operator and a ribosomal binding site upstream and a 6xHis-Tag downstream of the coding sequence were added. </p>
+
<html>
 +
<div class="container">
 +
  <div class="row">
 +
    <div class="col mx-2">
 +
 
 +
        <h3>Profile</h3>
 +
        <table style=“width:80%“>
 +
        <tr>
 +
        <td><b>Name</b></td>
 +
        <td>Sortase A7M </td>
 +
        </tr>
 +
        <tr>
 +
        <td><b>Base pairs</b></td>
 +
        <td>470</td>
 +
        </tr>
 +
        <tr>
 +
        <td><b>Molecular weigth</b></td>
 +
        <td>17.85&nbspkDa</td>
 +
        </tr>
 +
        <tr>
 +
        <td><b>Origin</b></td>
 +
        <td><i>Staphylococcus aureus</i>, synthetic</td>
 +
        </tr>
 +
        <tr>
 +
        <td><b>Parts</b></td>
 +
        <td>Basic part</td>
 +
        </tr>
 +
        <tr>
 +
        <td><b>Properties</b></td>
 +
        <td> calcium-independent, transpeptidase, linking sorting motif LPXTG to poly-glycine Tag </td>
 +
        </tr>
 +
        </table>
 +
<h3> Usage and Biology</h3>
 +
<p> Sortase&nbspA7M (SrtA7M) is a calcium-independent transpeptidase derived from the enzyme Sortase&nbspA from Staphylococcus aureus.<sup id="cite_ref-1" class="reference">
 +
  <a href="#cite_note-1">[1]</a>
 +
</sup> Similar to Sortase&nbspA, SrtA7M connects two peptides covalently, one with a poly-glycine (poly-G) at the N-terminus and one with LPXTG motif at the C-terminus. It acts by cleaving between Gly and Thr of the LPXTG motif and catalyzing the formation of an amide bond to the poly-G sequence.<sup id="cite_ref-2" class="reference">
 +
  <a href="#cite_note-2">[2] </a>
 +
</sup>
 +
 
 +
<h3>Methods</h3>
 +
<p>The basic part Sortase&nbspA7M is produced and purified as the composite part <a href="https://parts.igem.org/Part:BBa_K3187005"target="_blank">BBa_K3187005</a>(Sortase&nbspA7M).
 +
For the expression and purification of SrtA7M, a part containing T7 promoter, a lac operator and a ribosomal binding site <a href="https://parts.igem.org/Part:BBa_K3187029MMM"target="_blank">BBa_K3187029</a> N-terminal and a 6xHis-Tag with a TGA stop codon <a href="https://parts.igem.org/Part:BBa_K3187024"target="_blank">(BBa_K3187024)</a> C-terminal of the coding sequence were added.
 +
The production was performed in the <i>E.&nbsp;Coli</i> strain BL21 (DE3). The purification was done with the <a href="#"target="_blank">GE Healthcare ÄTKA Pure machine</a> which is a machine for FPLC. </p>
 +
 
 +
<h3>Results</h3>
 +
 
 +
 
 +
</div>
 +
</div>
 +
 
 +
</div>
 +
</html>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 12:33, 16 October 2019


Sortase A7M (Ca2+-independent variant)

Profile

Name Sortase A7M
Base pairs 470
Molecular weigth 17.85&nbspkDa
Origin Staphylococcus aureus, synthetic
Parts Basic part
Properties calcium-independent, transpeptidase, linking sorting motif LPXTG to poly-glycine Tag

Usage and Biology

Sortase&nbspA7M (SrtA7M) is a calcium-independent transpeptidase derived from the enzyme Sortase&nbspA from Staphylococcus aureus. [1] Similar to Sortase&nbspA, SrtA7M connects two peptides covalently, one with a poly-glycine (poly-G) at the N-terminus and one with LPXTG motif at the C-terminus. It acts by cleaving between Gly and Thr of the LPXTG motif and catalyzing the formation of an amide bond to the poly-G sequence. [2]

Methods

The basic part Sortase&nbspA7M is produced and purified as the composite part BBa_K3187005(Sortase&nbspA7M). For the expression and purification of SrtA7M, a part containing T7 promoter, a lac operator and a ribosomal binding site BBa_K3187029 N-terminal and a 6xHis-Tag with a TGA stop codon (BBa_K3187024) C-terminal of the coding sequence were added. The production was performed in the E. Coli strain BL21 (DE3). The purification was done with the GE Healthcare ÄTKA Pure machine which is a machine for FPLC.

Results

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 445
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]