Team:HokkaidoU Japan/Project

From 2011.igem.org

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==        Intro ==
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==Abstruct==
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   We further developed "Dr. ''E. coli''": our project of iGEM 2010. Last year, we showed that Type 3 Secretion System (T3SS) works in ''E. coli'' by injecting GFP into RK13 cells. We thought this system can be applied to direct reprogramming of somatic cells among many other things.
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   Bacteria living around us evolved ways to effect their surrounding environment. This is done by using secretion systems. There are six known types of secretion systems: one through six. These are called Type One Secretion System (T1SS), T2SS, T3SS and so on. Some system only secrete designated proteins outside, others can translocate proteins directly to other cells. Secretion systems able to inject proteins into other cell caught many researchers imagination.  
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   This year we repeated previous competitions experiment, one more time showing that GFP can be really injected into a target cell with it. And then, as a second step, we tested T3SS performance and tried to make it more convenient. For this purpose we designed a plasmid backbone which can instantly produce ready-to-inject fusion proteins from ordinary biobrick part. Using it, we tried to further characterize this system by injecting characteristic proteins.
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   Ours was no exception. We chose to use T3SS for our experiments. During iGEM 2010 we found that E.Coli with a part of Salmonella genome library expresses T3SS. This presented opportunity to work with the amazing machinery without involving pathogenic bacteria.  
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Results are still to come.
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   This year we repeated previous competitions experiment, one more time showing that GFP can be really injected into a target cell with it. And then continued to characterise it. An further develop data sheet of T3SS possibilities.
 
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==        Type Three Secretion System Data Sheet ==
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==Introduction to T3SS==
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   T3SS is a system of pathogenic gram-negative bacterium such as ''Salmonella'', ''Yersinia'' and EPEC (entero pathogenic ''E. coli''). Using this system bacteria can inject whole protein molecules through a syringe like organelle named T3S Apparatus. During iGEM 2010 we found that ''E. coli'' with a part of Salmonella genome library expresses T3SS. This presented opportunity to work with the amazing machinery without involving pathogenic bacteria.
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Still to come.
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[[Team:HokkaidoU_Japan/Project/T3SS|Read more]]
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==        Plasmid Backbone for protein injection ==
 
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==Plasmid Backbone for protein injection==
   We developed plasmid backbone which can attach tags needed for secretion and various other functions to a chosen protein biobrick. This can be used for big scale screening of various protein domains for their inject-ability.
   We developed plasmid backbone which can attach tags needed for secretion and various other functions to a chosen protein biobrick. This can be used for big scale screening of various protein domains for their inject-ability.
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More info still to come.
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[[Team:HokkaidoU_Japan/Project/Backbone|Read more]]
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==        Structure of Injectable Proteins ==
 
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==Investigation of T3SS-injectable proteins==
   Here we will discus the structure of proteins which are injected.  
   Here we will discus the structure of proteins which are injected.  
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Still to come.
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[[Team:HokkaidoU_Japan/Project/GSK|Read more]]
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{{:Team:HokkaidoU_Japan/footer}}
{{:Team:HokkaidoU_Japan/footer}}

Revision as of 07:40, 29 September 2011

HokkaidoUJapan

Contents

Abstruct

   We further developed "Dr. E. coli": our project of iGEM 2010. Last year, we showed that Type 3 Secretion System (T3SS) works in E. coli by injecting GFP into RK13 cells. We thought this system can be applied to direct reprogramming of somatic cells among many other things.

   This year we repeated previous competitions experiment, one more time showing that GFP can be really injected into a target cell with it. And then, as a second step, we tested T3SS performance and tried to make it more convenient. For this purpose we designed a plasmid backbone which can instantly produce ready-to-inject fusion proteins from ordinary biobrick part. Using it, we tried to further characterize this system by injecting characteristic proteins.

Results are still to come.


Introduction to T3SS

   T3SS is a system of pathogenic gram-negative bacterium such as Salmonella, Yersinia and EPEC (entero pathogenic E. coli). Using this system bacteria can inject whole protein molecules through a syringe like organelle named T3S Apparatus. During iGEM 2010 we found that E. coli with a part of Salmonella genome library expresses T3SS. This presented opportunity to work with the amazing machinery without involving pathogenic bacteria.

Read more


Plasmid Backbone for protein injection

   We developed plasmid backbone which can attach tags needed for secretion and various other functions to a chosen protein biobrick. This can be used for big scale screening of various protein domains for their inject-ability.

Read more


Investigation of T3SS-injectable proteins

   Here we will discus the structure of proteins which are injected.

Read more



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