中国兽医科学2024，Vol.54Issue(9)：1216-1223,8.DOI:10.16656/j.issn.1673-4696.2024.0155
非洲猪瘟病毒DP71L蛋白原核表达及其多克隆抗体制备
Prokaryotic expression of African swine fever virus DP71L protein and preparation of polyclonal antibody
摘要
Abstract
To study the biological function of DP71L protein of African swine fever virus(ASFV),the specific primers of ASFV CN/GS/2018 DP71L were designed according to the gene sequence of ASFV CN/GS/2018 DP71L.The DP71L gene was amplified and cloned into the prokaryotic expression vector pET28a(+),and then transformed into Escherichia coli BL21(DE3)to induce expression.BALB/c mice were immu-nized with the purified recombinant DP71L protein to prepare polyclonal antibody.The serum titer of the polyclonal antibody was determined by indirect ELISA,and the specificity of the polyclonal anti-body was identified by Western-blot.The results showed that the recombinant DP71L protein was ex-pressed in soluble form and inclusion body form.The titer of polyclonal antibody was 1∶64 000 measured by indirect ELISA.Western-blot results showed that the antibody specifically recognized the DP71L protein.The results showed that the recombinant DP71L protein was successfully expressed and the poly-clonal antibody was prepared,which laid a foundation for further study on the biological function of DP71L protein.关键词
非洲猪瘟/DP71L蛋白/原核表达/多克隆抗体Key words
African swine fever/DP71L protein/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
李志煜,马旭升,马永华,郑海学..非洲猪瘟病毒DP71L蛋白原核表达及其多克隆抗体制备[J].中国兽医科学,2024,54(9):1216-1223,8.基金项目
兰州市科技计划项目(2022-2-112,2021-RC-52) （2022-2-112,2021-RC-52）
甘肃省自然科学基金项目(18JR3RA167) （18JR3RA167）
甘肃农业大学基金项目(GAU-ZDKC-202216,GAU-XKJS-2018-073,GAU-KYQD-2017RCZX-11,GAU-QDFC-2020-11) （GAU-ZDKC-202216,GAU-XKJS-2018-073,GAU-KYQD-2017RCZX-11,GAU-QDFC-2020-11）
甘肃省科技厅国际合作项目(144WCGA169) （144WCGA169）
国家重点研发计划项目(2021YFD1800100) （2021YFD1800100）
非洲猪瘟亚单位疫苗研发项目(2021YF-D1801300) （2021YF-D1801300）
家畜重要疫病感染与免疫防控的基础理论研究项目(22ZD6NA001) （22ZD6NA001）
中央高校基本科研业务费专项(学科交叉创新团队建设项目)(lzujbky-2022-ct02) （学科交叉创新团队建设项目）
中国农业科学院创新计划项目(CAAS-CSLPDCP-2023002) （CAAS-CSLPDCP-2023002）
国家生猪技术创新中心项目(NCTIP-XD/C03) （NCTIP-XD/C03）
家畜重要疫病诊断研制与产业化项目(22ZD6NA012) （22ZD6NA012）
农业技术产业体系建设项目(CARS-35) （CARS-35）
"十四五"广东省揭榜挂帅项目(2023SDZG02) （2023SDZG02）
第三批甘肃省拔尖领军人才培养项目 （）
