中国动物检疫2024，Vol.41Issue(4)：103-110,8.DOI:10.3969/j.issn.1005-944X.2024.04.019
H9N2亚型禽流感病毒M1和NA基因在昆虫细胞中的表达
Expression of M1 and NA Genes of H9N2 Subtype Avian Influenza Virus in Insect Cells
摘要
Abstract
In order to express M1 and NA proteins of H9N2 subtype avian influenza virus(AIV)through insect cells and to identify their immune activity,M1 and NA genes were amplified by PCR,recombinant transfer vectors of pFastBacDual-M1 and pFastBacDual-NA were constructed based on pFastBacDual acting as a recombinant transfer vector;the positive recombinant transfer vectors were respectively transformed into DH10Bac competent cells to obtain recombinant bacmids of rBacmid-M1 and rBacmid-NA;Sf9 insect cells were transfected by the recombinant bacmids to obtain recombinant baculovirus containing M1 and NA genes,that were,rBV-M1 and rBV-NA;the expression of M1 and NA proteins was identified by IFA and Western-blot,and NA protein was used as a coated antigen to identify its reactivity by the indirect ELISA.The results showed that specific green fluorescence was observed by IFA identification,M1 and NA proteins were about 28 and 52 ku,respectively,as tested by Western-blot,and the expressed NA protein could specifically react with H9N2 positive serum.In conclusion,M1 and NA proteins with reactivity could be specifically expressed in insect cells.A foundation was laid for further development of diagnostic techniques and vaccine for H9N2 subtype AIV.关键词
禽流感病毒/H9N2亚型/M1蛋白/NA蛋白/Bac-to-Bac杆状病毒表达系统Key words
AIV/H9N2 subtype/M1 protein/NA protein/Bac-to-Bac baculovirus expression system分类
农业科技引用本文复制引用
王粲,张民秀,谢芝勋,李孟,罗思思,李丹,阮志华,谢丽基,谢志勤..H9N2亚型禽流感病毒M1和NA基因在昆虫细胞中的表达[J].中国动物检疫,2024,41(4):103-110,8.基金项目
广西重点研发计划项目(桂科AB21076004) （桂科AB21076004）
"广西八桂学者"项目(2019A50) （2019A50）
广西重大专项项目(桂科AA23062050) （桂科AA23062050）
广西科技先锋队项目(桂农科盟202409-01) （桂农科盟202409-01）
