| 猪瘟病毒E2蛋白酵母表达及其免疫活性研究 |
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| 引用本文： | 周稳,金前跃,陈晓,丁培阳,李旭峰,王垚,柴永笑,张雨航,周恩民,郭军庆,郅玉宝,张改平.猪瘟病毒E2蛋白酵母表达及其免疫活性研究[J].动物医学进展,2020(1):1-7. |
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| 作者姓名： | 周稳 金前跃 陈晓 丁培阳 李旭峰 王垚 柴永笑 张雨航 周恩民 郭军庆 郅玉宝 张改平 |
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| 作者单位： | 西北农林科技大学动物医学院;河南省农业科学院动物免疫学重点实验室/河南省动物免疫学重点实验室/农业部动物免疫学重点实验室;河南农业大学牧医工程学院 |
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| 基金项目： | 国家重点研发计划项目(2017YFD0501103) |
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| 摘 要： | 猪瘟(CSF)是由猪瘟病毒(CSFV)引起的一种急性、热性、高度接触性传染病,呈世界性分布,对养猪业危害严重。猪瘟病毒属于黄病毒科、瘟病毒属,在自然条件下只感染猪。E2蛋白是猪瘟病毒的重要结构蛋白,也是猪瘟病毒的主要保护性抗原。以猪瘟病毒E2蛋白胞外区基因为模板,构建毕赤酵母表达载体pPICZαA-E2,并通过电转获得毕赤酵母X-33阳性克隆,经SDS-PAGE、Western blot鉴定筛选出猪瘟病毒E2蛋白表达菌株。通过亲和层析技术纯化蛋白,并将蛋白免疫动物,进行免疫原性分析。结果表明,猪瘟病毒E2蛋白在毕赤酵母中成功分泌性表达,经纯化纯度达95%以上;动物试验显示,该蛋白具有良好的免疫原性,为猪瘟病毒亚单位疫苗及相关检测技术的开发奠定了基础。
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| 关 键 词： | 猪瘟病毒 E2蛋白 毕赤酵母 蛋白纯化 免疫活性 |
Expression of Classical Swine Fever Virus E2 Protein in Pichia pastoris and Study on Immunological Activity |
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| Institution: | (College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi,712100,China;Key Laboratory of Animal Immunology,Henan Academy of Agricultural Science/Henan Provincial Key Laboratory of Animal Immunology/Key Laboratory of Animal Immunology of Ministry of Agricultural,Zhengzhou,Henan,450002,China;College of Animal Science and Veterinary Medicine,Henan Agricultural University,Zhengzhou,Henan,450002,China) |
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| Abstract: | Classical swine fever(CSF) is an acute,hot,highly contagious infectious disease caused by classical swine fever virus(CSFV),which has a worldwide distribution and is serious harmful to the pig industry.CSFV belongs toPestivirus genus of Flaviviridae family,the virus in natural conditions only infects pigs.E2 protein is an important structural protein of CSFV and it is the main protective antigen of CSFV.In this study,pPICZαA-E2 vector was constructed using the E2 protein extracellular domain gene of CSFV as the template.The X-33 positive clones were obtained by electroporation,and the E2 protein expression strain was identified by SDS-PAGE and Western blot analysis.E2 protein was purified by affinity chromatography,and the purified protein was immunized against animals for immunogenicity analysis.The results showed that the E2 protein of CSFV was successfully expressed in X-33 and reached to 95% purity.Animal experiments showed that the E2 protein has high immunological activity.This study laid a foundation for the development of subunit vaccine and related detection technology of CSFV. |
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| Keywords: | Classical swine fever virus E2 protein Pichia pastoris protein purification immunological activity |
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