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Chimeric Newcastle disease virus nucleocapsid with parts of viral hemagglutinin-neuraminidase and fusion proteins

机译:嵌合的新城疫病毒核衣壳,带有部分病毒血凝素神经氨酸酶和融合蛋白

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摘要

The nucleocapsid (NP) protein of Newcastle disease virus (NDV) self-assembled in Escherichia coli as ring-like and herringbone-like particles. Several chimeric NP proteins were constructed in which the antigenic regions of the hemagglutinin-neuraminidase (HN) and fusion (F) proteins of NDV, myc epitope, and six histidines (a hexa-His tag) were linked to the C-terminus of the NP monomer. These chimeric proteins were expressed efficiently in soluble form in E. coli as detected by Western blot analysis. Electron microscopy of the purified products revealed that they self-assembled into ring-like particles. These chimeric particles exhibited antigenicity of the myc epitope, suggesting that the foreign sequences were exposed on the surface of the particles. Chickens inoculated with the chimeric particles mounted an immune response against NDV, suggesting the possibility of use of the ring-like particle as a carrier of immunogens in subunit vaccines and immunological reagents.
机译:新城疫病毒(NDV)的核衣壳(NP)蛋白在大肠杆菌中自组装成环状和人字形颗粒。构建了几种嵌合NP蛋白,其中NDV的血凝素神经氨酸酶(HN)和融合蛋白(F),myc表位和六个组氨酸(hexa-His标签)的抗原区连接到NP的C端。 NP单体。如通过蛋白质印迹分析所检测,这些嵌合蛋白在大肠杆菌中以可溶形式有效表达。纯化产物的电子显微镜显示,它们自组装成环状颗粒。这些嵌合颗粒表现出myc表位的抗原性,表明外源序列暴露在颗粒表面上。接种了嵌合颗粒的鸡对NDV产生了免疫应答,这表明有可能使用环状颗粒作为亚单位疫苗和免疫试剂中免疫原的载体。

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