首页|非洲猪瘟病毒DP71L蛋白原核表达及其多克隆抗体制备

非洲猪瘟病毒DP71L蛋白原核表达及其多克隆抗体制备

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  • 国家科技期刊平台
  • NETL
  • NSTL
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为深入研究非洲猪瘟病毒(ASFV)DP71L蛋白的生物学功能,根据非洲猪瘟病毒ASFV CN/GS/2018 DP71L基因序列设计特异性引物,通过扩增DP71L基因并克隆到原核表达载体pET28a(+)后转入大肠杆菌BL21(DE3)诱导表达,通过SDS-PAGE、Western-blot进行鉴定,并以纯化出的重组DP71L蛋白免疫BALB/c小鼠,制备多克隆抗体,间接ELISA测定多克隆抗体效价,Western-blot鉴定多克隆抗体的特异性.结果显示,重组DP71L蛋白以可溶物和包涵体两种形式表达;间接ELISA测得多克隆抗体血清效价为1∶64 000;Western-blot结果显示,该抗体能特异性识别DP71L蛋白.结论:成功地表达了DP71L重组蛋白,制备了多克隆抗体,为深入研究DP71L蛋白生物学功能奠定了基础.
Prokaryotic expression of African swine fever virus DP71L protein and preparation of polyclonal antibody
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.

African swine feverDP71L proteinprokaryotic expressionpolyclonal antibody

李志煜、马旭升、马永华、郑海学

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甘肃农业大学动物医学院,甘肃 兰州 730070

中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室,甘肃兰州 730000

非洲猪瘟 DP71L蛋白 原核表达 多克隆抗体

兰州市科技计划项目兰州市科技计划项目甘肃省自然科学基金项目甘肃农业大学基金项目甘肃农业大学基金项目甘肃农业大学基金项目甘肃农业大学基金项目甘肃省科技厅国际合作项目国家重点研发计划项目非洲猪瘟亚单位疫苗研发项目家畜重要疫病感染与免疫防控的基础理论研究项目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中国农业科学院创新计划项目国家生猪技术创新中心项目家畜重要疫病诊断研制与产业化项目农业技术产业体系建设项目"十四五"广东省揭榜挂帅项目第三批甘肃省拔尖领军人才培养项目

2022-2-1122021-RC-5218JR3RA167GAU-ZDKC-202216GAU-XKJS-2018-073GAU-KYQD-2017RCZX-11GAU-QDFC-2020-11144WCGA1692021YFD18001002021YF-D180130022ZD6NA001lzujbky-2022-ct02CAAS-CSLPDCP-2023002NCTIP-XD/C0322ZD6NA012CARS-352023SDZG02

2024

10.16656/j.issn.1673-4696.2024.0155

中国兽医科学
中国农业科学院兰州兽医研究所

中国兽医科学

CSTPCD北大核心
影响因子：0.524
ISSN：1673-4696
年,卷(期)：2024.54(9)