Difference between revisions of "Part:BBa K1769000"

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Dimeric Hrs FYVE is a small binding molecule that can bind to phosphatidylinositol 3-phosphate (PtdIns(3)P), a receptor that exists on endosome. Since it is more stable then monomeric FYVE, it is widely used to recognize and locate PtdIns(3)P.
 
Dimeric Hrs FYVE is a small binding molecule that can bind to phosphatidylinositol 3-phosphate (PtdIns(3)P), a receptor that exists on endosome. Since it is more stable then monomeric FYVE, it is widely used to recognize and locate PtdIns(3)P.
  
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===Usage and Biology===
 
===Usage and Biology===
 
Many pathogen, such as <i>Plasmodium falciparum<i> that causes malaria and  
 
Many pathogen, such as <i>Plasmodium falciparum<i> that causes malaria and  
 
<i>Phytophthora<i> spp. that cause a wide range of plant disaeses,inhibits the immune system of host cell by secreting effector proteins. These effector protein translocate into host cell by binding to a outer membrane receptor, PI3P. This biobrick can not only serve as a useful tool to study the signal transduction pathway of PI3P but can also be a weapon to stop these pathogen's infection.  
 
<i>Phytophthora<i> spp. that cause a wide range of plant disaeses,inhibits the immune system of host cell by secreting effector proteins. These effector protein translocate into host cell by binding to a outer membrane receptor, PI3P. This biobrick can not only serve as a useful tool to study the signal transduction pathway of PI3P but can also be a weapon to stop these pathogen's infection.  
  
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== '''Introduction''' ==
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To stop the effector protein secreted by P.infestans from binding to PI3P receptor and translocate into the cell, we designed a competitive inhibitor called FYVE protein domain. This protein domain was obtained from Hrs gene from mice. However, a monomeric FYVE protein domain is not stable in plant cell and the affinity for PI3P is not high enough to compete with effector protien, Avr3, as you can see in figure1. Therefore we designed a linker region to connect the two protein domain together.(Fig2)
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[[File:Nymu-Fyve-mono_mod.jpg|left|thumb|300px|'''Fig1 monomeric FYVE and Avr3 competition model''']]
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[[[[File:Nymu-Fyve-dimer_mod.jpg|thumb|300px|'''Fig dimeric FYVE and Avr3 competition model''']]
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 09:43, 17 September 2015

Dimeric FYVE

Dimeric Hrs FYVE is a small binding molecule that can bind to phosphatidylinositol 3-phosphate (PtdIns(3)P), a receptor that exists on endosome. Since it is more stable then monomeric FYVE, it is widely used to recognize and locate PtdIns(3)P.

Usage and Biology

Many pathogen, such as Plasmodium falciparum<i> that causes malaria and <i>Phytophthora<i> spp. that cause a wide range of plant disaeses,inhibits the immune system of host cell by secreting effector proteins. These effector protein translocate into host cell by binding to a outer membrane receptor, PI3P. This biobrick can not only serve as a useful tool to study the signal transduction pathway of PI3P but can also be a weapon to stop these pathogen's infection.


Introduction

To stop the effector protein secreted by P.infestans from binding to PI3P receptor and translocate into the cell, we designed a competitive inhibitor called FYVE protein domain. This protein domain was obtained from Hrs gene from mice. However, a monomeric FYVE protein domain is not stable in plant cell and the affinity for PI3P is not high enough to compete with effector protien, Avr3, as you can see in figure1. Therefore we designed a linker region to connect the two protein domain together.(Fig2)

File:Nymu-Fyve-mono mod.jpg
Fig1 monomeric FYVE and Avr3 competition model
[[
File:Nymu-Fyve-dimer mod.jpg
Fig dimeric FYVE and Avr3 competition model

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 110
    Illegal BglII site found at 329
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]