Difference between revisions of "Part:BBa K3718002"

Latest revision as of 06:37, 17 October 2021

Customizable detection and expression with increased growth

Part II of the Toolkit for Detection with Proliferation Control

The Modular Receptor Platform (MRP) is designed according to the paper published in 2018 (Chang et al, 2018). When the antigen/ligand binds to the nanobody, dimerization of the protein CadC domains occur. As a result, this allows the binding of the pCadBA operon region and transcription downstream is promoted at the same time.

The VHH region has high flexibility as it can be customized for binding with different targets, so that any specific targets with single-domain antibodies available will be able to bind with the VHH region for detection. Users can add their own VHH region after the transmembrane region DNA via restriction ligation using KasI. For more details about the design of the customizable VHH region, please visit the registry of the part BBa_K3718001.

For the developers to clone the gene(s) that they want to express, a multiple cloning site (MCS) K660500 to allow for the insertion of the target genes. arcA and iclR genes are originally knocked out using Part I of our toolkit to reduce growth. When the antigen is detected, arcA and iclR are expressed to restore growth while protein(s) of choice are expressed.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal prefix found in sequence at 67
Illegal suffix found in sequence at 629
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 67
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal SpeI site found at 630
Illegal PstI site found at 644
Illegal NotI site found at 73
Illegal NotI site found at 637
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 67
Illegal BglII site found at 2337
Illegal BamHI site found at 912
Illegal XhoI site found at 906
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found in sequence at 67
Illegal suffix found in sequence at 630
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found in sequence at 67
Illegal XbaI site found at 82
Illegal SpeI site found at 630
Illegal PstI site found at 644
1000
COMPATIBLE WITH RFC[1000]

Reference

H. Waegeman, J. Beauprez, H. Moens, J. Maertens, M. De Mey, M. R. Foulquié-Moreno, J. J. Heijnen, D. Charlier, and W. Soetaert, “Effect of ICLR and ARCA knockouts on biomass formation and metabolic fluxes in escherichia coli K12 and its implications on understanding the metabolism of escherichia coli BL21 (DE3),” BMC Microbiology, vol. 11, no. 1, p. 70, 2011.

H.-J. Chang, P. Mayonove, A. Zavala, A. De Visch, P. Minard, M. Cohen-Gonsaud, and J. Bonnet, “A modular receptor platform to expand the sensing repertoire of bacteria,” ACS Synthetic Biology, vol. 7, no. 1, pp. 166–175, 2017.

@@ Line 5: / Line 5: @@
-Part II of toolkit for Detection with Proliferation Control
+<p>Part II of the Toolkit for Detection with Proliferation Control</p>
-The Modular Receptor Platform (MRP) is designed according to the paper published in 2018 (Chang et al, 2018). When the antigen/ligand binds to the nanobody, dimerization of the protein CadC domains occur. As a result, this allows the binding of the pCadBA operon region and transcription downstream is promoted at the same time. The VHH region has high flexibility as it can be customized for binding with different targets, so that any specific targets with single domain antibodies available will be able to bind with the VHH region for detection. Users can add their own VHH region after the transmembrane region DNA via restriction ligation.
+<p>The Modular Receptor Platform (MRP) is designed according to the paper published in 2018 (Chang et al, 2018). When the antigen/ligand binds to the nanobody, dimerization of the protein CadC domains occur. As a result, this allows the binding of the pCadBA operon region and transcription downstream is promoted at the same time.</p>
-For the developers to clone the genes that they want to express, a multiple cloning site (MCS) K660500 is present in the same region as the genes that allow for restored growth so it will be expressed when the antigen is detected. The MCS allows for the insertion of the target genes to be less complex and be less time consuming.
+https://2021.igem.org/wiki/images/thumb/6/63/T--Hong_Kong_UCCKE--MRP.png/800px-T--Hong_Kong_UCCKE--MRP.png
+<p>The VHH region has high flexibility as it can be customized for binding with different targets, so that any specific targets with single-domain antibodies available will be able to bind with the VHH region for detection. Users can add their own VHH region after the transmembrane region DNA via restriction ligation using KasI. For more details about the design of the customizable VHH region, please visit the registry of the part BBa_K3718001.</p>
+https://2021.igem.org/wiki/images/thumb/4/42/T--Hong_Kong_UCCKE--using_toolkit.jpeg/800px-T--Hong_Kong_UCCKE--using_toolkit.jpeg
+<p>For the developers to clone the gene(s) that they want to express, a multiple cloning site (MCS) K660500 to allow for the insertion of the target genes. <i>arcA</i> and <i>iclR</i> genes are originally knocked out using Part I of our toolkit to reduce growth. When the antigen is detected, <i>arcA</i> and <i>iclR</i> are expressed to restore growth while protein(s) of choice are expressed.</p>
@@ Line 24: / Line 30: @@
 <partinfo>BBa_K3718002 parameters</partinfo>
 <!-- -->
+<h3>Reference</h3>
+<p> H. Waegeman, J. Beauprez, H. Moens, J. Maertens, M. De Mey, M. R. Foulquié-Moreno, J. J. Heijnen, D. Charlier, and W. Soetaert, “Effect of ICLR and ARCA knockouts on biomass formation and metabolic fluxes in escherichia coli K12 and its implications on understanding the metabolism of escherichia coli BL21 (DE3),” <i>BMC Microbiology</i>, vol. 11, no. 1, p. 70, 2011.</p>
+<p>H.-J. Chang, P. Mayonove, A. Zavala, A. De Visch, P. Minard, M. Cohen-Gonsaud, and J. Bonnet, “A modular receptor platform to expand the sensing repertoire of bacteria,” <i>ACS Synthetic Biology</i>, vol. 7, no. 1, pp. 166–175, 2017.</p>