Difference between revisions of "Part:BBa K2549053"

 
 
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<partinfo>BBa_K2549053 short</partinfo>
 
<partinfo>BBa_K2549053 short</partinfo>
  
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This part is a linker that contains four glycines and one serine. Small size glycines enable the fusion protein to be flexible and meanwhile polar serine residues ensure solubility. Copies of G4S linker could be changed to achieve optimal flexibility or separation of the adjacent domains.
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This part is a linker that contains four glycines and one serine. Small size glycines enable the fusion protein to be flexible and meanwhile polar serine residues ensure solubility. Copies of G4S linker could be changed to achieve optimal flexibility or separation of the adjacent domains.  We include this part as it is required for the parts collection [[Part:BBa_K2549016]] ~ [[Part:BBa_K2549043]].
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==Literature Characterization by AFCM-Egypt==
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The study used G4S Linker (E7O2V) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. The Engineered Cell Line was provided by the Lohmueller Lab, University of Pittsburgh. To express flow cytometry for testing of the G4S Linker.
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<html><div align="center"style="border:solid #17252A; width:50%;float:center;"><img style="                              max-width:850px;
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width:75%;
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height:auto;
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position: relative;
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top: 50%;
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left: 35%;
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transform: translate( -50%);
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padding-bottom:25px;
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padding-top:25px;
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"src="https://static.igem.wiki/teams/4586/wiki/literature-characterisation-parts/capture.png">
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<p class=MsoNormal align=center style='text-align:left;border:none;width:98% ;justify-content:center;'><span
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lang=EN style='font-size:11.0pt;line-height:115%'>The study used express flow cytometry to assess live Jurkat cells, which appear blue (negative), and engineered Jurkat cells, which appear green (positive). To express an scFv-based anti-CD19 CAR containing a G4S linker.
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</span></p></div></html>
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<br><br><br><br>
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<html><div align="center"style="border:solid #17252A; width:50%;float:center;"><img style="                              max-width:850px;
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width:75%;
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height:auto;
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position: relative;
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top: 50%;
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left: 35%;
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transform: translate( -50%);
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padding-bottom:25px;
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padding-top:25px;
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"src="https://static.igem.wiki/teams/4586/wiki/literature-characterisation-parts/g4s-linker.png">
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<p class=MsoNormal align=center style='text-align:left;border:none;width:98% ;justify-content:center;'><span
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lang=EN style='font-size:11.0pt;line-height:115%'>Flow cytometry was used to watch the cell binding of GLPB30 antibodies to CAR T-cell constructs containing G4S-linked ScFvs. (A) By using an anti-human PE secondary antibody after an anti-G4S GLPB30 antibody, they could find 3–23/B3 CAR-transgene expression. Two days after CD3+ pan-T cell mRNA transduction, staining was carried out. (B) N-terminal MYC tags could be found on anti-CD19 CAR with an (G4S)3
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linker, and they could be observed on lentiviral-transfected pan-T cells. (c) In contrast to MYC-negative cells, MYC-positive CAR T cells showed higher GLPB30 staining.
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</span></p></div></html>
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 19:50, 11 October 2023


G4S linker

This part is a linker that contains four glycines and one serine. Small size glycines enable the fusion protein to be flexible and meanwhile polar serine residues ensure solubility. Copies of G4S linker could be changed to achieve optimal flexibility or separation of the adjacent domains. We include this part as it is required for the parts collection Part:BBa_K2549016 ~ Part:BBa_K2549043.

Literature Characterization by AFCM-Egypt

The study used G4S Linker (E7O2V) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. The Engineered Cell Line was provided by the Lohmueller Lab, University of Pittsburgh. To express flow cytometry for testing of the G4S Linker.

The study used express flow cytometry to assess live Jurkat cells, which appear blue (negative), and engineered Jurkat cells, which appear green (positive). To express an scFv-based anti-CD19 CAR containing a G4S linker.





Flow cytometry was used to watch the cell binding of GLPB30 antibodies to CAR T-cell constructs containing G4S-linked ScFvs. (A) By using an anti-human PE secondary antibody after an anti-G4S GLPB30 antibody, they could find 3–23/B3 CAR-transgene expression. Two days after CD3+ pan-T cell mRNA transduction, staining was carried out. (B) N-terminal MYC tags could be found on anti-CD19 CAR with an (G4S)3 linker, and they could be observed on lentiviral-transfected pan-T cells. (c) In contrast to MYC-negative cells, MYC-positive CAR T cells showed higher GLPB30 staining.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]