Composite

Part:BBa_K3064024

Designed by: Hanxiao Feng   Group: iGEM19_NUDT_CHINA   (2019-09-28)
Revision as of 17:55, 29 September 2019 by Petersberg (Talk | contribs) (→‎Special Design)


mmuGlucagon-GGGGS-mmulgG-Fc-P2A-HA-mmuTrim21

This composite part is made up of six basic parts:mmuGlucagon,GGGGS,mmuIgG-Fc,P2A,HA and mmuTrim21.
It includes the parts we need to construct a GCGR degradation system. mmuGlucagon can bing with glucagon receptor specifically and it plays a role as a navigator in the composite part. GGGGS Linker is mostly used as a linker for protein and it is one of the best fold breaking linker to expose fusion protein partner well.Immunoglobulin G (IgG) is a type of antibody, which binds many kinds of pathogens and protects the body from infection. Also, it can be recognized and bound with TRIM21,an E3 ubiquitin-protein ligase that has been applied to a protein degradation method known as Trim-Away.The bicistronic P2A peptide is a kind of self-cleaved sequence, which breaks when translated. It can be used to construct the polycis vector and realize the co-translation of two proteins under the control of the same promoter.The HA-tag we use in our system has the length of 27 bps. It is a really useful tool to link antibody and Trim21.Trim21, as referred,can trigger the degradation of certain protein.
Composed of the basic parts above, this composite part can degrade GCGR specifically and thus lower glucose content to realize our goal.

Usage and Biology

We use this composite part to degrade the content of glucagon-receptor specifically which can lead to the decrease of glucose content in mice and cause the change of hepatic metabolism. By observing the changes we can further explore the relationship between two symptoms.

Special Design

Special designs were taken to optimize the applicability and adaptive of such parts. GGGGS is one of the best fold breaking linker to expose fusion protein partner well. With it we can construct a composite part with better stability and applicability. Two sequences of proteins could be fused with their corresponding fragments at the same time and it can dramatically simplify the cloning process. Special_design_of_composite_part.png <B/r> Figure 1.Representation of the function of the composite part. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 800
    Illegal BamHI site found at 1503
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 901
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 125

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