Difference between revisions of "Part:BBa K5301024"
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<partinfo>BBa_K5301024 short</partinfo>
 
<partinfo>BBa_K5301024 short</partinfo>
  
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===Usage and Biology===
 
In order to produce large nanodiscs more conveniently, we hope to flexibly extend the length of MSP according to demand, and thus propose the concept of multi-polymer MSP, which refers to large circular MSPs through end-to-end connections of multiple MSP fragments.  
 
In order to produce large nanodiscs more conveniently, we hope to flexibly extend the length of MSP according to demand, and thus propose the concept of multi-polymer MSP, which refers to large circular MSPs through end-to-end connections of multiple MSP fragments.  
 
We used NW15 as the basic MSP and selected three types of linkers (Spy/Sdy/Snoop) to achieve the connection of different MSP fragments through the formation of covalent bonds, and adopted rigorous design to prevent self-cyclization of each fragment of the multi-polymer MSP. Finally, the successful cyclization of large circular MSPs is characterized by the fluorescence of mCherry after the combination of mCherry [1-10] and mCherry [11].
 
We used NW15 as the basic MSP and selected three types of linkers (Spy/Sdy/Snoop) to achieve the connection of different MSP fragments through the formation of covalent bonds, and adopted rigorous design to prevent self-cyclization of each fragment of the multi-polymer MSP. Finally, the successful cyclization of large circular MSPs is characterized by the fluorescence of mCherry after the combination of mCherry [1-10] and mCherry [11].
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<p>We also utilized AlphaFold 2 to simulate the overall structure of multi-polymerized MSP, and obtained multi-polymerized MSP with the correct conformation, as shown in the figure. </p>
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<div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:min-content;"><div style="zoom:0.3;overflow:hidden;">
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https://static.igem.wiki/teams/5301/parts/msp1.gif
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</div><div class="thumbcaption">
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Figure 1.Alpha Fold 2 prediction model of the overall structure of multi-polymerized MSP
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</div></div></div></div>
  
 
===Sequence and Features===
 
===Sequence and Features===

Revision as of 13:53, 1 October 2024


Multi-polymer MSP, which refers to large circular MSPs through end-to-end connections of multiple MS

Usage and Biology

In order to produce large nanodiscs more conveniently, we hope to flexibly extend the length of MSP according to demand, and thus propose the concept of multi-polymer MSP, which refers to large circular MSPs through end-to-end connections of multiple MSP fragments. We used NW15 as the basic MSP and selected three types of linkers (Spy/Sdy/Snoop) to achieve the connection of different MSP fragments through the formation of covalent bonds, and adopted rigorous design to prevent self-cyclization of each fragment of the multi-polymer MSP. Finally, the successful cyclization of large circular MSPs is characterized by the fluorescence of mCherry after the combination of mCherry [1-10] and mCherry [11].

We also utilized AlphaFold 2 to simulate the overall structure of multi-polymerized MSP, and obtained multi-polymerized MSP with the correct conformation, as shown in the figure.

msp1.gif

Figure 1.Alpha Fold 2 prediction model of the overall structure of multi-polymerized MSP

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 3141
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 3141
    Illegal NotI site found at 1720
    Illegal NotI site found at 4038
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1239
    Illegal BglII site found at 2323
    Illegal BglII site found at 2695
    Illegal BglII site found at 3557
    Illegal BglII site found at 3929
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 3141
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 3141
    Illegal AgeI site found at 1035
    Illegal AgeI site found at 1828
    Illegal AgeI site found at 2220
    Illegal AgeI site found at 2980
  • 1000
    COMPATIBLE WITH RFC[1000]