Part:BBa_K4768005

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

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Molecular Modeling

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Characterisation

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Conclusion and Perspectives

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References

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