农业生物技术学报2024，Vol.32Issue(10)：2391-2401,11.DOI:10.3969/j.issn.1674-7968.2024.10.017
乙型脑炎病毒囊膜蛋白I176R位点突变株的生物学特性比较研究
Comparative Study on the Biological Characteristics of Japanese encephalitis virus Envelope Protein I176R Site Mutant Strain
摘要
Abstract
Japanese encephalitis virus(JEV)is a serious zoonotic virus,and the envelope protein(E)plays a key role in the neurovirulence of JEV.Previous studies have found that the mutation of EI176R site of recombinant Japanese encephalitis virus(rJEV-EI176R)can weaken its neurovirulence.In order to identify the biological characteristics of rJEV-EI176R,the E gene sequence of rJEV-EI176R was cloned and analyzed,and the bioinformatics characteristics of E protein were analyzed.The growth curve of rJEV-EI176R,the adsorption difference of various cells and the inflammatory level of mouse microglia cell line(BV-2)induced by rJEV-EI176R were determined by qPCR,plaque assay and indirect immunofluorescence assay(IFA).The results showed that the positive charge residue of E protein of rJEV-EI176R increased by 1,the isoelectric point increased by 0.37,and the instability index increased by 0.39.The number and proportion of α-helix,β-sheet,extended strand and random coil of E protein changed,and the amino acid at E176 site had different connections with nearby amino acids.The growth curve of rJEV-EI176R on BV-2 cells did not change significantly and could infect nerve cells and renal cells from different sources.The adsorption capacity of rJEV-EI176R was significantly improved(P＜0.05).After inoculation of BV-2 cells with rJEV-EI176R,the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interferon-γ-induced protein-10(IP-10)were significantly lower than those of the parental strain at 12 and 24 h(P＜0.05).This study enriched the biological characteristics of E protein of JEV strain,and provides basic data for further study of JEV attenuated mechanism and vaccine development.关键词
乙型脑炎病毒(JEV)/囊膜蛋白(E)/I176R位点突变/神经毒力致弱/生物学特性Key words
Japanese encephalitis virus(JEV)/Envelope protein(E)/I176R mutation/Neurovirulence attenuation/Biological characteristics分类
农业科技引用本文复制引用
唐冕耀,曹三杰,赵勤,石艳萍,曹玉琴,文翼平,伍锐,杜森焱,黄小波,颜其贵..乙型脑炎病毒囊膜蛋白I176R位点突变株的生物学特性比较研究[J].农业生物技术学报,2024,32(10):2391-2401,11.基金项目
国家自然科学基金(31972722) （31972722）
