Coding

Part:BBa_K5526008

Designed by: SIYUAN WU   Group: iGEM24_HWFLA-Beijing   (2024-09-02)

Anti-PD-L1

Anti-PD-L1


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 25
    Illegal AgeI site found at 364
  • 1000
    COMPATIBLE WITH RFC[1000]

Anti-PD-L1 (BBa_K5526008) Documentation

Part: BBa_K5526008 (Anti-PD-L1)

Profile

Name: Anti-PD-L1

Base Pairs: 372bp

Origin: Cell, synthesized

Properties: Anti-PD-L1 is an immune checkpoint inhibitor that blocks the recognition of PD-L1 expressed by tumor cells with T cells, enhancing the immune system's effect on tumors. It has been widely used in the clinical treatment of various types of cancer.

Usage and Biology

PD-L1 is a protein expressed on the surface of many tumor cells that allows them to evade the immune system by binding to the PD-1 receptor on T cells. By blocking the interaction between PD-L1 and PD-1, Anti-PD-L1 reactivates the T cell-mediated immune response, helping the immune system recognize and destroy tumor cells. It has shown remarkable therapeutic effects, particularly in the field of immunotherapy.

Gene map of Anti-PD-L1
Figure 1. Gene map of Anti-PD-L1

Cultivation, Purification, and SDS-PAGE

AntiPD-L1 is a set gene that encodes an immunomodulatory protein, which activates the immune system to kill tumor cells. pUC57-mini is the plasmid skeleton. We applied PCR to amplify the antiPD-L1 gene (372bp) and pUC57-plldr (3150bp), and used agarose gel electrophoresis to confirm the PCR products. The result is that antiPD-L1 has a length of 372bp, as shown in Figure 2.

PCR production identification by agarose gel electrophoresis
Figure 2. The identification of PCR production by agarose gel electrophoresis. Left: The graph shows that plactate 1 has a length of 3150 bp. Right: The graph shows that antiPD-L1 has a length of 372bp.

After optimizing the protein expression conditions, the constructed strains were expanded for culture. 5mM lactic acid was used to induce tumor drug expression in EcN1917. The experimental results are shown in Figure 3. The size of the Anti-PD-L1 protein is about 24kDa, and a large amount of target protein was successfully expressed.

Detection of Anti-PD-L1 protein expression by SDS-PAGE
Figure 3. Detection of Anti-PD-L1 protein expression by SDS-PAGE

Future studies will focus on investigating the expression and tumor inhibition effects of the constructed probiotics in a tumor environment.

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