Fluorescent leishmania plasmid, macrophage-targeting sPD-1 lizard leishmania expression system and preparation method

By designing the fluorescent Leishmania plasmid PLN-mCherry and sPD-1 genes, the simultaneous expression of fluorescent protein and sPD-1 protein in Leishmania lizard was achieved and the targeting of macrophages was solved, which was not possible in the prior art at the same time The issue of expression and recognition provides new cancer treatments.

CN119242679APending Publication Date: 2025-01-03WEIFANG MEDICAL UNIV
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Patent Information

Application Number
CN202411329161.5
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-09-24
Publication Date
2025-01-03

AI Technical Summary

Technical Problem

The existing Leishmania plasmids cannot express fluorescent protein and sPD-1 protein simultaneously, and cannot specifically recognize and target macrophages, limiting their application in cancer treatment.

Method used

A fluorescent Leishmania plasmid PLN-mCherry was designed to bind to the sPD-1 gene and electrotransfect the lizard Leishmania to achieve the simultaneous expression of fluorescent protein and sPD-1 protein in any Leishmania, and The sPD-1 protein was expressed by targeting macrophages and analyzed for its induced cytokine changes.

Benefits of technology

The PLN-mCherry plasmid and sPD-1 lizard Leishmania expression system was successfully constructed, and the simultaneous expression of fluorescent proteins and sPD-1 proteins was achieved, targeting macrophages, and providing new cancer treatment ideas and methods.

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Abstract

The invention belongs to the technical field of lizard leishmania expression, and particularly relates to a fluorescent leishmania plasmid, a macrophage-targeting sPD-1 lizard leishmania expression system and a preparation method. The plasmid is formed by connecting a fragment 1 and a fragment 2 end to end, wherein the nucleotide sequence of the fragment 1 is as shown in SEQ ID NO: 1; the nucleotide sequence of the fragment 2 is as shown in SEQ ID NO: 2; the plasmid is PLN-mCherry, and the nucleotide sequence of the plasmid is as shown in SEQ ID NO: 3. A host bacterium of the expression system is lizard leishmania and contains a recombinant plasmid NC-sPD-1, and the recombinant plasmid NC-sPD-1 is composed of a plasmid described in any one of claims 1-2 and an sPD-1 sequence; according to the present invention, the PLN-mCherry leishmania expression plasmid is successfully constructed, and the cherry red fluorescence is expressed in the lizard leishmania; the lizard leishmania expression system expressing sPD-1 is obtained, fluorescent protein and target protein can be expressed in any leishmania, and expression of the two proteins does not affect each other.
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