Difference between revisions of "Part:BBa K2833008"
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<h1>Design</h1>
 
<h1>Design</h1>
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We got the INP from (<html><a href='https://parts.igem.org/Part:BBa_K523013'>BBa_K523013</a></html>) constructed by iGEM11_Edinburgh. To test the surface display efficiency of INP, we add YFP to the downstream of INP, and expressed the fusion protein under the control of J23104 and B0032.
+
We got the INP from (<html><a href='https://parts.igem.org/Part:BBa_K523013'>BBa_K523013</a></html>) constructed by iGEM11_Edinburgh. To test the surface display efficiency of INP, we add YFP to the downstream of INP, and expressed the fusion protein under the control of (<html><a href='https://parts.igem.org/Part:BBa_J23104'>J23104</a></html>) and (<html><a href='https://parts.igem.org/Part:BBa_B0032'>B0032</a></html>) .
  
 
<h1>Microscopy result</h1>
 
<h1>Microscopy result</h1>

Revision as of 12:34, 8 October 2018


J23104+B0032+INP+YFP

Ice nucleation protein(INP)is a secretory outer membrane protein from Pseudomomas syringae P. flurorescens and several other Gram—negative bacteria.In the bacterial surface display technology，INP is widely used as a functional carrier protein.

Design

We got the INP from (BBa_K523013) constructed by iGEM11_Edinburgh. To test the surface display efficiency of INP, we add YFP to the downstream of INP, and expressed the fusion protein under the control of (J23104) and (B0032) .

Microscopy result

We observed the fluorescence under the super-resolution microscope. As figure 1 shows, The intracellular expression of sfGFP displayed the rode-shape of E.coli, while the signal of surface displayed YFP showed the dotted pattern, which suggested that the YFP were gathered and distributed at the surface of E.coli..

super-resolution microscopy of INP-YFP left: intracellular expression of GFP; Right: surface display of YFP via INP


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 467
  • 1000
    COMPATIBLE WITH RFC[1000]