Difference between revisions of "Part:BBa K4768005"
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<partinfo>BBa_K4768005 parameters</partinfo>
 
<partinfo>BBa_K4768005 parameters</partinfo>
 
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<h2>Introduction</h2>
 
<h2>Introduction</h2>
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            <figcaption class="normal"><span class="titre-image"><i><b>Figure 1: T7Nterm-SL-Pertuzumab structure.</b></i></span></figcaption>
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<p>The CALIPSO part BBa_K4768005 is composed of the N-terminal subunit of the T7 RNA polymerase (residues 1 to 180) fused to the anti-HER2 antibody Pertuzumab through a soluble linker. This gene is under transcriptional control of an SP6 promoter and T7 terminator.</p>
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<p>This part, coupled to the part <a href="https://parts.igem.org/Part:BBa_K4768006" target="blank">BBa_K4768006</a> containing the C-terminal subunit of the T7 RNA polymerase, has been designed to develop a split T7 RNAP-based biosensor capable of recognizing HER-2, an epidermal growth factor that is overexpressed in cancer cells [1], in solution.</p>
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<p>The HER2-induced T7 RNAP complex was designed from two existing constructs: a split T7 RNAP-based biosensor for the detection of rapamycin [2] and a split luciferase conjugated with antibodies capable of recognizing HER2 [3]. We decided to merge the relevant functionalities of these two constructs and created a new biosensor that transduces HER2 binding to gene expression activation. </p>
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            <figcaption class="normal"><span class="titre-image"><i><b>Figure 2: Recognition of HER2 extracellular domain induces functional assembly of the split T7 RNA polymerase, which enables gene expression of target gene under control of a T7 promoter.</b></i></span></figcaption>
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<h2>Construction</h2>
 
<h2>Construction</h2>

Revision as of 09:41, 8 October 2023


split T7 RNA polymerase (Nterm) conjugated to Pertuzumab with a soluble linker

Part for Expression of the split T7 RNA polymerase (Nterm) conjugated to Pertuzumab with a soluble linker in PURE System

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 40
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 636
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 40
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 40
    Illegal AgeI site found at 1104
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 21
    Illegal SapI.rc site found at 1740



Introduction

Figure 1: T7Nterm-SL-Pertuzumab structure.

The CALIPSO part BBa_K4768005 is composed of the N-terminal subunit of the T7 RNA polymerase (residues 1 to 180) fused to the anti-HER2 antibody Pertuzumab through a soluble linker. This gene is under transcriptional control of an SP6 promoter and T7 terminator.

This part, coupled to the part BBa_K4768006 containing the C-terminal subunit of the T7 RNA polymerase, has been designed to develop a split T7 RNAP-based biosensor capable of recognizing HER-2, an epidermal growth factor that is overexpressed in cancer cells [1], in solution.

The HER2-induced T7 RNAP complex was designed from two existing constructs: a split T7 RNAP-based biosensor for the detection of rapamycin [2] and a split luciferase conjugated with antibodies capable of recognizing HER2 [3]. We decided to merge the relevant functionalities of these two constructs and created a new biosensor that transduces HER2 binding to gene expression activation.

Figure 2: Recognition of HER2 extracellular domain induces functional assembly of the split T7 RNA polymerase, which enables gene expression of target gene under control of a T7 promoter.

Construction

TXXXXXX.

Molecular Modeling

TXXXXXX.

Characterisation

TXXXXXX.

Conclusion and Perspectives

TXXXXXX.

References

  1. article 1 xxxxxxxx
  2. article 2 xxxxxxx
    3.