非洲猪瘟病毒E199L-CP204L-E183L串联序列在杆状病毒表达系统中的表达及免疫效果评价OA北大核心CSTPCD

In this study,African swine fever virus tandem sequence of E199L,CP204L and E183L were cloned into pFastBac1 to obtain recombinant plasmids pFastBac-CMV-EGFP-P2A-E199L-CP204L-E183L-HA.The recombinant plasmids were transformed into DH10Bac competent cells.Following antibiotic resis-tance and blue-white screening,and PCR verification,recombinant bacmids were obtained.Recombinant baculoviruses rBac-E199L-CP204L-E183L were obtained by transfecting sf9 cells with recombinant bacmids.Western-blot and indirect immunofluorescence assay(IFA)were used to detect protein expres-sion of recombinant baculoviruses in sf9 cells.The results showed that IFA detected obvious fluores-cence and Western-blot detected bands matching the size of the target proteins,indicating that the constructed recombinant baculoviruses were successfully expressed.Subsequently,the immunogenicity of rBac-E199L-CP204L-E183L was investigated by detecting the serum specific antibodies and T lympho-cyte proliferation.The results showed that antibodies against pE199L,p30,and p54 were detected on 14 days post second immunization,respectively.The proliferation of T lymphocyte of the immunized mice is significantly higher than that of control mice.Overall,three tandem proteins were successfully ex-pressed in baculovirus expression system and immunization of the recombinant baculovirus induced hu-moral immune responses,and stimulated lymphocyte proliferation.