尼帕病毒N蛋白在昆虫细胞上的表达及蛋白生物信息学分析  

Expression and Bioinformatics Analysis of Nipah Virus N Protein in Insect Cells

作　　者：王腾芬 叶玲玲 李书宁 张伟 胡世享 卓娜 艾军 陈培富[1] WANG Tengfen;YE Lingling;LI Shuning;ZHANG Wei;HU Shixiang;ZHUO Na;AI Jun;CHEN Peifu(Yunnan Agricultural University,Kunming 650201,China;Kunming Customs Technology Center,Kunming 651000,China;Xishuangbanna Animal Disease Prevention and Control Center,Jinghong 666199,China)

机构地区：[1]云南农业大学,云南昆明650201 [2]昆明海关技术中心,云南昆明651000 [3]西双版纳州动物疫病预防控制中心,云南景洪666199

出　　处：《华北农学报》2024年第S1期259-266,共8页Acta Agriculturae Boreali-Sinica

基　　金：重大野生动物疫病跨境传播机制及监测技术研究(2022YFC2601605)。

摘　　要：为获得具有和天然蛋白质类似功能与活性的尼帕病毒核衣壳蛋白(N蛋白),通过PCR扩增NiV N蛋白开放阅读框基因,克隆pFastBac^(TM)HTB载体,转化DH10Bac感受态细胞,转染sf9昆虫细胞,蛋白免疫印迹鉴定,生物信息学分析研究N蛋白特性。结果表明,经PCR和双酶切鉴定,获得重组质粒pFastBac^(TM)HTB-NiV-N;经SDS-PAGE和Western Blotting鉴定,获得具有免疫活性的重组蛋白Bacmid-NiV-N;经分子生物信息学软件分析预测,重组蛋白含有532个氨基酸,分子大小为58.34 ku,为不稳定、亲水性蛋白,无跨膜区,蛋白二级结构主要以无规则卷曲为主,有5个T细胞抗原表位和18个B细胞抗原表位。上述研究为深入进行NiV N蛋白的功能和建立相应抗原检测方法提供物质基础。In order to obtain Nipah virus nucleocapsid protein(N protein)with similar function and activity to natural proteins,the NiV N protein open reading frame gene was amplified by PCR,cloned into pFastBac^(TM)HTB vector,transformed into DH10Bac receptive cells,and transfected into sf9 insect cells.The protein was identified by Western Blotting and bioinformatics analysis was used to study the characteristics of N protein.The results showed that the recombinant plasmid pFastBacMHTB-NiV-N was obtained through PCR and double enzyme digestion iden-tification.Recombinant protein Bacmid-NiV-N with immune activity was identified by SDS-PAGE and Western Blot-ting.According to molecular bioinformatics software analysis and prediction,the recombinant protein contained 532 amino acids with a molecular size of 58.34 ku.It was an unstable and hydrophilic protein without a transmembrane region.The secondary structure of the protein was mainly irregular curls,with 5 T cell epitopes and 18 B cell epitopes.The above research provides a material basis for further understanding the function of NiV N protein and establishing corresponding antigen detection methods.

关 键 词：尼帕病毒 N蛋白 昆虫细胞 生物信息学分析 

分 类 号：S432.4[农业科学—植物病理学] Q78[农业科学—农业昆虫与害虫防治]