中国兽医科学2024，Vol.54Issue(11)：1458-1464,7.DOI:10.16656/j.issn.1673-4696.2024.0183
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
Expression of African swine fever virus E199L-CP204L-E183L in baculovirus expression system and assessment of its immunogenicity
摘要
Abstract
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.关键词
非洲猪瘟病毒/E199L/CP204L/E183L/杆状病毒/免疫评价Key words
African swine fever virus/E199L/CP204L/E183L/baculovirus/immunological evaluation分类
农业科技引用本文复制引用
张寅寅,李智晟,王晨晓,倪洁,刘静宜,陈鸿军,孙耀贵..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[J].中国兽医科学,2024,54(11):1458-1464,7.基金项目
国家重点研发计划项目(2021YFD1801401) （2021YFD1801401）
