Difference between revisions of "Part:BBa K1995005"
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<partinfo>BBa_K1995005 short</partinfo>
 
<partinfo>BBa_K1995005 short</partinfo>
  
 
Based on BBa_K1995004 (BC), we added INP-N and GFP to the upstream of BC. Using INP-N, we anchored GFP and BC out of the outer membrane of E.coli in order to increase the binding efficiency and using GFP to detect the concentration of cupric and mercuric ions.
 
Based on BBa_K1995004 (BC), we added INP-N and GFP to the upstream of BC. Using INP-N, we anchored GFP and BC out of the outer membrane of E.coli in order to increase the binding efficiency and using GFP to detect the concentration of cupric and mercuric ions.
  
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'''Ice nucleation protein (INP)'''
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==Ice nucleation protein (INP)==
 
   
 
   
 
We used INP to anchor GFP and BC out of the outer membrane of E. coli, firstly increasing the probability of the meeting of BC and heavy metal ions and secondly using the fluorescence quenching of GFP to detect the concentration of heavy metal ions.
 
We used INP to anchor GFP and BC out of the outer membrane of E. coli, firstly increasing the probability of the meeting of BC and heavy metal ions and secondly using the fluorescence quenching of GFP to detect the concentration of heavy metal ions.
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[[File:INP.png]]
 
[[File:INP.png]]
  
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'''GFP reporter'''
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==GFP reporter==
  
 
GFP is an excellent reporter in synthetic biology. We added GFP into IC for following functions:
 
GFP is an excellent reporter in synthetic biology. We added GFP into IC for following functions:
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[[File:0053.png]]
 
[[File:0053.png]]
  
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'''In different strains of E. coli'''
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==In different strains of E. coli==
  
 
We tested the binding efficiency of Hg2+ with IC using E. coli and engineering E.coli. (Fig. 4)
 
We tested the binding efficiency of Hg2+ with IC using E. coli and engineering E.coli. (Fig. 4)
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[[File:0054.png]]
 
[[File:0054.png]]
  
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'''Binding heavy metals in fish gut'''
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==Binding heavy metals in fish gut==
  
 
We detect the binding efficiency of IC in fish gut by feeding fishes with IC metal catcher and get the following chart. (Fig. 5)
 
We detect the binding efficiency of IC in fish gut by feeding fishes with IC metal catcher and get the following chart. (Fig. 5)
  
 
[[File:0055.png]]
 
[[File:0055.png]]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 13:15, 16 October 2016


IC (INP-N-GFP-C)

Based on BBa_K1995004 (BC), we added INP-N and GFP to the upstream of BC. Using INP-N, we anchored GFP and BC out of the outer membrane of E.coli in order to increase the binding efficiency and using GFP to detect the concentration of cupric and mercuric ions.

Ice nucleation protein (INP)

We used INP to anchor GFP and BC out of the outer membrane of E. coli, firstly increasing the probability of the meeting of BC and heavy metal ions and secondly using the fluorescence quenching of GFP to detect the concentration of heavy metal ions.

Ice nucleation protein (INP) is an excretion surface protein. It is widely used for establishing germ surface display system. It contains an internal repeated domain (IRD) and an N-terminal anchoring domain (fig.1). Studies have shown that decrease the time of repetition of IRD can also get a better-off rate of displaying. So we decreased the time of repetition of IRD to reduce pressure of synthesis of E. coli.

INP.png

GFP reporter

GFP is an excellent reporter in synthetic biology. We added GFP into IC for following functions:

① Initial survey of the expression of IC.

② Detect the binding efficiency and other features of IC.

③ Detect and measure the concentration of cupric and mercuric ions.

0052.png

0053.png

In different strains of E. coli

We tested the binding efficiency of Hg2+ with IC using E. coli and engineering E.coli. (Fig. 4)

0054.png

Binding heavy metals in fish gut

We detect the binding efficiency of IC in fish gut by feeding fishes with IC metal catcher and get the following chart. (Fig. 5)

0055.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 429
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1181
    Illegal SapI.rc site found at 238