首页|H9N2禽流感病毒HA蛋白mRNA疫苗的构建表达及鉴定

H9N2禽流感病毒HA蛋白mRNA疫苗的构建表达及鉴定

扫码查看
原文链接
  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
H9N2 亚型禽流感病毒(AIV)是严重危害家禽养殖业的病原,由于目前的疫苗无法应对突发的AIV 疫情,因此本研究设计一种基于血凝素(HA)基因的抗H9N2 亚型禽流感病毒mRNA 疫苗.在原有Puc57 克隆载体基础上,构建mRNA体外转录模板,将PCR得到的HA基因和EGFP基因插入模板中构建重组质粒Puc57-HA和Puc57-EGFP.将重组质粒线性化后,进行体外转录、加帽、加尾、纯化等一系列修饰并转染鸡成纤维细胞(DF-1),观察 48 h荧光表达情况,并用Western-blot检测HA蛋白在细胞中的表达.结果显示,48 h出现绿色荧光,Western-blot显示HA蛋白成功表达.本试验为H9N2 禽流感mRNA疫苗的研制奠定了基础.
Construction and expression and identification of H9N2 avian influenza virus HA protein mRNA vaccine
H9N2 avian influenza virus is one of the etiological agents that seriously threaten the poultry industry.As the current vaccine can not cope with the sudden AIV epidemic,this study designed an mRNA vaccine against H9N2 subtype avian influenza virus based on hemagglutinin(HA)gene.On the ba-sis of the original Puc57 cloning vector,mRNA transcription template was constructed in vitro,and the HA gene and EGFP gene obtained by PCR were inserted into the template to construct the recombinant plas-mid Puc57-HA and Puc57-EGFP.The recombinant plasmid was linearized,transcribed in vitro,capped,cau-dated and purified,and then transfected into chicken fibroblasts(DF-1)cells,fluorescence expression was observed for 48 h,and the expression of HA protein in cells was detected by Western-blot.The re-sults showed that green fluorescence appeared at 48 h,and Western-blot showed that HA protein was suc-cessfully expressed.This experiment laid a foundation for the development of mRNA vaccine for H9N2 avian influenza.

avian influenza virusmRNA vaccinein vitro transcription

花悦、宋奇珊、王东东、杨思敏、王颖、任建乐、牛胜、尚桂军、赵宇军、田文霞

展开 >

山西农业大学动物医学学院,山西 晋中 030801

山西高等创新研究院功能蛋白结构解析山西省重点实验室,山西 太原 030032

禽流感 mRNA疫苗 体外转录

中美疾病预防控制中心合作项目中美疾病预防控制中心合作项目中美疾病预防控制中心合作项目山西省重点研发计划山西省现代农业产业技术体系建设专项山西省现代农业产业技术体系建设专项山西省现代农业产业技术体系建设专项山西省科技创新青年人才团队项目功能蛋白结构解析山西省重点实验室项目山西省"1331工程"建设项目

5U01IP001106-04-002022HX116E2900901-022021021305010012021-072022-072023CYJSTX15-0920220405100102220210401091000620211331-13

2024

10.16656/j.issn.1673-4696.2024.0089

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(5)
  • 17