Difference between revisions of "Team:HokkaidoU Japan/Aggregation"

 
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<br>  When you harvest <span style="font-style: italic">E.coli</span> on your laboratory, is there the way to aggregate <span style="font-style: italic">E.coli</span> efficiently with the SAP?  We try to suggest the expression of the SAP on <span style="font-style: italic">E. coli</span> flagellum. Perhaps, a new method for the <span style="font-style: italic">E.coli</span> aggregation, except for Ag43 which has been adopted by some iGEM teams[1][2], may be found.
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<br>  When you harvest <span style="font-style: italic">E. coli</span> on your laboratory, is there a efficiently way to aggregate <span style="font-style: italic">E. coli</span> with the SAP?  We would like to try to suggest a possibility of the expression of <span style="font-style: italic">E. coli</span> aggregation by expression of the SAP fused to <span style="font-style: italic">E. coli</span> flagellum. Perhaps, a new method for the <span style="font-style: italic">E. coli</span> aggregation, except for Ag43 which has been adopted by some iGEM teams[1][2], may be found.
  
 
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<td style="border-style: none"; align="center"><h2>Fig. 1. Image of aggregation using SAP
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<td style="border-style: none"; align="center"><span class="small">Fig. 1. Image of aggregation using SAP
<br><span style="font-style: italic">E.coli</span> are aggregated by the binding through SAP. SAP is expressed on the fllagerum.</h2></td>
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<br><span style="font-style: italic">E. coli</span> are aggregated by the binding through SAP. SAP is expressed on the fllagerum.</span></td>
 
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<br>FliC is an important domain which forms the filament (Fig. 2)[3]. It is known that any protein can be inserted into its domain without losing the function of the flagellar. So we think that the aggregation of <span style="font-style: italic">E.coli</span> because of the bindings between the flagellar is likely to happen by inserting SAP, for example, RADA16-I and P<span class="sitatuki">11</span>-4. Since it has been reported that the overexpression of the Ag43 decreases the survival rate of E.coli, we have to search for an alternative method for the aggregation of <span style="font-style: italic">E.coli</span>.   
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<br>FliC is an important domain which forms the filament (Fig. 2)[3]. It is known that any protein can be inserted into its domain without losing the function of the flagella. So we think that the aggregation of <span style="font-style: italic">E. coli</span> because of the bindings between the flagella is likely to happen by inserting SAP, for example, RADA16-I and P<span class="sitatuki">11</span>-4. Since it has been reported that the overexpression of the Ag43 decreases the survival rate of <span style="font-style: italic">E. coli</span>, we have to search for an alternative method for the aggregation of <span style="font-style: italic">E. coli</span>.   
<br>Therefore we propose that inserting the SAP has an advantage of improving the efficiency of the consecutive chemical reactions when <span style="font-style: italic">E.coli</span> which have different functions approach each other.
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<br>Therefore we propose that inserting the SAP has an advantage of improving the efficiency of the consecutive chemical reactions when <span style="font-style: italic">E. coli</span> which have different functions approach each other.
 
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<td style="border-style: none"; align="center"><h2>Fig. 2. Image of <span style="font-style: italic">E.coli</span>'s firament  
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<td style="border-style: none"; align="center"><span class="small">Fig. 2. Image of filament of <span style="font-style: italic">E. coli</span>   
<br>Any protein can be inserted into the FliC.</h2></td>
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<br>Any protein can be inserted into the FliC.</span></td>
 
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<br>  There may still be room for improvement for the financial aspect or the efficiency regarding the way to harvest <span style="font-style: italic">E.coli</span> in the laboratory. And a more simple method would be much more desirable without any expensive biological device. At last we wish that the research will be more conducted of the difference between the efficiency of the aggregation of <span style="font-style: italic">E.coli</span> with the biological tools, for example Ag43, RADA16-I, and P<span class="sitatuki">11</span>-4. And we also desire for the possibility of the aggregation of <span style="font-style: italic">E.coli</span> with the SAP we suggest.
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<br>  If the method for industrial production of self-assembling peptides by using <span style="font-style: italic">E. coli </span> is established, it will contribute to disperse the utilization of hydrogel. To achieve this, we should focus on the synthesis and transportation of self-assembling peptides. We mentioned about “Transportation” above. Furthermore, we hope that other teams make further developments regarding “Synthesis”.And also the aggregation methods for <span style="font-style: italic">E. coli </span> in lab may be improved in the aspects of the cost or efficiency. From this aspects, the simpler methods without utilizing Ag43, His- tag, or RADA16-I are desirable. We expect that the understanding about the differences of the aggregating efficiency among the methods utilizing these proteins will be expanded. In addition, we hope that the aggregation method for <span style="font-style: italic">E. coli </span> by utilizing the self-assembling peptides we proposed will be useful in the future.
 
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[1] <a href="https://2012.igem.org/Team:HokkaidoU_Japan/Project/Aggregation">HokkaidoU Japan 2012</a>
 
[1] <a href="https://2012.igem.org/Team:HokkaidoU_Japan/Project/Aggregation">HokkaidoU Japan 2012</a>
 
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[2] <a href"https://2014.igem.org/Team:Aberdeen_Scotland/Parts">Aberdeen Scotland 2014</a>
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[2] <a href="https://2014.igem.org/Team:Aberdeen_Scotland/Parts">Aberdeen Scotland 2014</a>
<br>[3] Kuwajima, G. 1988. Construction of a minimum-size functional flagellin of Escherichia coli. J. Bacteriol. 170:3305–3309.
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<br>[3] Kuwajima, G. 1988. Construction of a minimum-size functional flagellin of Escherichia coli. J. Bacteriol. 170:3305-3309.
  
 
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Latest revision as of 20:17, 19 October 2016

Team:HokkaidoU Japan - 2016.igem.org

 

Team:HokkaidoU Japan

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overview

When you harvest E. coli on your laboratory, is there a efficiently way to aggregate E. coli with the SAP? We would like to try to suggest a possibility of the expression of E. coli aggregation by expression of the SAP fused to E. coli flagellum. Perhaps, a new method for the E. coli aggregation, except for Ag43 which has been adopted by some iGEM teams[1][2], may be found.

aggregation
Fig. 1. Image of aggregation using SAP
E. coli are aggregated by the binding through SAP. SAP is expressed on the fllagerum.





design

FliC is an important domain which forms the filament (Fig. 2)[3]. It is known that any protein can be inserted into its domain without losing the function of the flagella. So we think that the aggregation of E. coli because of the bindings between the flagella is likely to happen by inserting SAP, for example, RADA16-I and P11-4. Since it has been reported that the overexpression of the Ag43 decreases the survival rate of E. coli, we have to search for an alternative method for the aggregation of E. coli.
Therefore we propose that inserting the SAP has an advantage of improving the efficiency of the consecutive chemical reactions when E. coli which have different functions approach each other.

filament
Fig. 2. Image of filament of E. coli
Any protein can be inserted into the FliC.





conclusion

If the method for industrial production of self-assembling peptides by using E. coli is established, it will contribute to disperse the utilization of hydrogel. To achieve this, we should focus on the synthesis and transportation of self-assembling peptides. We mentioned about “Transportation” above. Furthermore, we hope that other teams make further developments regarding “Synthesis”.And also the aggregation methods for E. coli in lab may be improved in the aspects of the cost or efficiency. From this aspects, the simpler methods without utilizing Ag43, His- tag, or RADA16-I are desirable. We expect that the understanding about the differences of the aggregating efficiency among the methods utilizing these proteins will be expanded. In addition, we hope that the aggregation method for E. coli by utilizing the self-assembling peptides we proposed will be useful in the future.


reference

[1] HokkaidoU Japan 2012
[2] Aberdeen Scotland 2014
[3] Kuwajima, G. 1988. Construction of a minimum-size functional flagellin of Escherichia coli. J. Bacteriol. 170:3305-3309.