中国兽医科学2024，Vol.54Issue(5)：609-614,6.DOI:10.16656/j.issn.1673-4696.2024.0089
H9N2禽流感病毒HA蛋白mRNA疫苗的构建表达及鉴定
Construction and expression and identification of H9N2 avian influenza virus HA protein mRNA vaccine
摘要
Abstract
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.关键词
禽流感/mRNA疫苗/体外转录Key words
avian influenza virus/mRNA vaccine/in vitro transcription分类
农业科技引用本文复制引用
花悦,田文霞,宋奇珊,王东东,杨思敏,王颖,任建乐,牛胜,尚桂军,赵宇军..H9N2禽流感病毒HA蛋白mRNA疫苗的构建表达及鉴定[J].中国兽医科学,2024,54(5):609-614,6.基金项目
中美疾病预防控制中心合作项目(5U01IP001106-04-00,2022HX116,E2900901-02) （5U01IP001106-04-00,2022HX116,E2900901-02）
山西省重点研发项目(202102130501001) （202102130501001）
山西省现代农业产业技术体系建设专项资金(2021-07,2022-07,2023CYJSTX15-09) （2021-07,2022-07,2023CYJSTX15-09）
山西省科技创新青年人才团队项目(202204051001022) （202204051001022）
功能蛋白结构解析山西省重点实验室资助项目(202104010910006) （202104010910006）
山西省1331工程项目(20211331-13) （20211331-13）
