Difference between revisions of "Part:BBa K2833009"
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<partinfo>BBa_K2833009 short</partinfo>
 
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Ice nucleation protein(INP)is a secretory outer membrane protein from Pseudomomas syringae，P． flurorescens and several other Gram—negative bacteria．Using its characteristic of forming ice nucleation at relatively warm temperatures(一2～一4‘C)，INP is originally applied in biological refrigeration field．In the bacterial surface display technology，INP is widely used as a functional carrier protein．Compared with other carrier proteins，INP has advantages such as stably expressing foreign proteins and showing the larger molecular weight proteins．INP cell—sur— face display technology has been developed to be a promising approach in many biotechn0109ical fields，such as whole cell bioeatalysts，whole cell bioadsorbence and biodegradation of environmental pollutants.  
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Ice nucleation protein(INP)is a secretory outer membrane protein from Pseudomomas syringae <I>P. flurorescens</I> and several other Gram—negative bacteria.In the bacterial <B>surface display</B> technology，INP is widely used as a functional carrier protein.
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<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 added sfGFP 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>.
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<h1>Microscopy result</h1>
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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 sfGFP showed the dotted pattern, which suggested that the sfGFP were gathered and distributed at the surface of <I>E.coli.</I>.
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[[File:T--BJRS China--009.jpg|thumbnail|center|500px|<b>Fig.1 super-resolution microscopy of INP-sfGFP</b> Left: intracellular expression of GFP; Right: surface display of GFP via INP]]<br><br>
  
 
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Revision as of 12:45, 8 October 2018


J23104+B0032+INP+sfGFP

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 added sfGFP 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 sfGFP showed the dotted pattern, which suggested that the sfGFP were gathered and distributed at the surface of E.coli..

Fig.1 super-resolution microscopy of INP-sfGFP Left: intracellular expression of GFP; Right: surface display of GFP 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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1050