Construction method and application of African swine fever virus protein eukaryotic expression vector with immunosuppressive effect

By constructing the eukaryotic expression vector pcDNA3.1-B119L of the African swine fever virus B119L gene, the inhibition of the cellular immune response was solved, and the problem of difficult inhibition of cells' immune response to Poly(dT:dT), Poly(I:C), and exogenous IFN-β in the prior art was solved, which proved the immunosuppressive function of the B119L protein and laid the foundation for related research.

CN119639816APending Publication Date: 2025-03-18YANGZHOU UNIV
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Patent Information

Application Number
CN202411879349.7
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-12-19
Publication Date
2025-03-18

AI Technical Summary

Technical Problem

The prior art is difficult to effectively inhibit the immune response of cells to Poly(dT:dT), Poly(I:C), and exogenous IFN-β, especially in the study of the B119L protein of African swine fever virus.

Method used

By designing specific primers and Flag tag sequences, the eukaryotic expression vector pcDNA3.1-B119L of the African swine fever virus B119L gene was constructed, and the expression of B119L protein was achieved through PCR amplification and enzyme cleavage and ligation to inhibit the immune response of cells to Poly(dT:dT), Poly(I:C), and exogenous IFN-β.

Benefits of technology

It successfully inhibited the immune response of cells to Poly(dT:dT), Poly(I:C), and exogenous IFN-β, proved the immunosuppressive function of B119L protein, and laid the foundation for the study of the biological function of B119L protein of African swine fever virus.

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Abstract

The invention discloses a construction method and application of an African swine fever virus protein eukaryotic expression vector with an immunosuppressive effect, and relates to the technical field of virus coding protein expression and function research biology. According to the eukaryotic expression vector, an African swine fever virus genome is taken as a template, a specific primer is adopted for PCR (Polymerase Chain Reaction) amplification, a PCR fragment is recovered and then is connected with a pcDNA3.1 vector to obtain a positive recombinant plasmid, namely the eukaryotic expression vector with the function of inhibiting stimulated immune response of cells to Poly (dT: dT), Poly (I: C) and exogenous IFN-beta. The construction method is simple and reasonable, after transfecting cell expression, the eukaryotic expression vector can be applied to inhibition of Poly (dT: dT), Poly (I: C) and exogenous IFN-beta stimulated cellular immune response, and establishment of an ASFV detection method and participation in vaccine development as a candidate target are facilitated.
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