中国兽医科学2025，Vol.55Issue(6)：740-747,8.DOI:10.16656/j.issn.1673-4696.2025.0093
非洲猪瘟病毒p15蛋白在昆虫细胞中的表达及其免疫原性评价
Expression and immunogenicity evaluation of African swine fever virus p15 protein in insect baculovirus expression system
摘要
Abstract
This study is dedicated to preparing the p15 protein of African swine fever virus(ASFV)by insect baculovirus expression system and evaluating its immunogenicity,thereby providing an exper-imental foundation for researching subunit vaccines against African swine fever.Initially,gene se-quence encoding the p15 protein was cloned into pFastBac1 vector to construct the recombinant plasmid pFastBac-p15.Subsequently,the plasmid was homologous to DH10Bac and transfected into insect cells to harvest recombinant baculovirus rBac-p15.The rBac-p15 was then used to infect sf9 cells for the ex-pression of the recombinant protein.Next,the target protein was purified through affinity chromatog-raphy and identified using SDS-PAGE,Western-blot,and IFA to assess the expression and reactivity of the recombinant protein.Finally,Mice were then immunized with p15 protein to evaluate the levels of humoral and cellular immune responses.The results demonstrated that the p15 protein was highly ex-pressed in the infected sf9 cells and could be specifically recognized by the convalescent pig serum of ASFV.The immunization of mice with the p15 protein induced the generation of high levels of specific anti-bodies and cytokines,which was conducive to stimulating the proliferation of CD4+and CD8+T lymphocytes in the spleen.In conclusion,this study successfully prepared the p15 protein using the insect bac-ulovirus expression system.This protein is capable of simultaneously inducing excellent humoral and cel-lular immune responses,which is conducive to the further research of African swine fever vaccines.关键词
非洲猪瘟病毒/p15蛋白/昆虫杆状病毒表达系统/免疫原性评价Key words
African swine fever virus/p15 protein/baculovirus expression system/immunogenicity evaluation分类
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党海迷,韩盛楠,伍国强,郑海学,杨洋,茹毅,郝荣增,蒋成辉,赵陇和,卢炳州,李亚军,胡馨予..非洲猪瘟病毒p15蛋白在昆虫细胞中的表达及其免疫原性评价[J].中国兽医科学,2025,55(6):740-747,8.基金项目
中央级公益性科研院所基本科研业务费专项(CAAS-ZDRW202409) （CAAS-ZDRW202409）
国家生猪产业技术体系项目(CARS-35) （CARS-35）
中央高校基本科研业务费专项(lzujbky-2022-ct02) （lzujbky-2022-ct02）
甘肃省自然科学基金重点项目(23YFNA0011) （23YFNA0011）
甘肃省科技重大专项(23ZDK A0002) （23ZDK A0002）
兰州市青年科技人才创新项目(2023-QN-3) （2023-QN-3）
