中国畜牧兽医2016，Vol.43Issue(10)：2762-2767,6.DOI:10.16431/j.cnki.1671-7236.2016.10.037
H3N2亚型猪流感病毒HA1、HA2基因的原核表达及抗原性分析
Prokaryotic Expression and Antigenic Analysis of H3N2 Swine Influenza Virus HA1 and HA2 Genes
摘要
Abstract
Hemagglutinin protein plays an important role in disease prevention and treatment of the swine influenza virus (SIV).In order to clone and express subtype H3N2 SIV A/swine/Henan/1/2010 (H3N2)HA1 and HA2 genes with chicken embryo allantoic fluid containing the H3N2 SIV after extraction of RNA.The HA1 gene and HA2 gene were amplified from the total RNA using RT-PCR and they were inserted into prokaryotic expression vector pET-28a (+)to construct recombinant expression vector.Then the vector was transformed and expressed in E .coli BL21 (DE3)pLyS.Then the bacteria were induced by IPTG and their lysates were ana-lyzed by SDS-PAGE and Western blotting,respectively.The monoclonal antibody 1C10,which was specific to HA protein,was used as primary antibody in Western blotting analysis.It was found that the expressed recombinant HA1 protein was 34.8 ku and recombinant HA2 protein was 23.2 ku analyzed by SDS-PAGE.As demonstrated by Western blotting,this HA1 expressed products showed the capacity of reacting with monoclonal antibody 1C10.All the results sugges-ted that the expressed HA1 and HA2 proteins of H3N2 influenza virus would be very helpful in the development of rapid diagnosis method and vaccine development,which would facilitate fur-ther study on the function of HA at the same time.关键词
H3N2亚型猪流感病毒/HA1基因/HA2基因/原核表达Key words
influenza A virus subtype H3N2/HA1 gene/HA2 gene/prokaryotic expression分类
农业科技引用本文复制引用
宋玉慧,王翔宇,王洁琼,潘梦梦,张剑,刘琳,李新生..H3N2亚型猪流感病毒HA1、HA2基因的原核表达及抗原性分析[J].中国畜牧兽医,2016,43(10):2762-2767,6.基金项目
河南省科技创新人才计划(144100510009)；郑州市前沿技术研究开发项目(141PQYJS566)；河南省产学研合作项目(15210700096) （144100510009）
