作 者：
单春辉;李金斗;吴佳奇;冯嘉轩;丁佳欣;陈凯楠;郭春红;丁壮
关键词：
鹅细小病毒病;鹅副黏病毒病;VP3基因;嵌合型VLPs
摘 要：
基于新城疫病毒样颗粒(Newcastle disease virus-like particles,NDV VLPs)载体平台,采用胞外域替换的策略、昆虫杆状病毒表达系统、蔗糖密度梯度离心和透射电镜等方法,构建嵌合型病毒样颗粒.将鹅细小病毒(goose par-vovirus,GPV)的VP3基因与鹅副黏病毒病(goose paramyxovirus)的HN基因相连并命名为rVP3并构建了表达rVP3蛋白的重组杆状病毒rBV-rVP3,将其与本实验室前期构建的rBV-M和rBV-HN共感染sf9细胞72 h后获得鹅细小病毒病-鹅副黏病毒病二联嵌合型病毒样颗粒(GPV-NDV cVLPs).采用超速离心和蔗糖密度梯度离心的方法纯化GPV-NDV cVLPs,并通过透射电镜进行观察,可见与野生NDV形态相似的cVLPs;利用Western blot技术对GPV-NDV cVLPs各组分蛋白进行鉴定,结果显示蛋白表达均正确.本试验为预防鹅细小病毒病(goose parvovir-us infection,GP)和鹅副黏病毒病提供一种安全的、可"一针多防"的新型疫苗候选株.
译 名：
Construction and identification of chimeric virus-like particles of goose parvovirus and goose paramyxovirus
关键词：
goose parvovirus infection%goose paramyxovirus%VP3 gene%biochimeric VLPs
摘 要：
To provide a new green,safe and"one-shot"vaccine candidate for the prevention of goose parvovirus infection(GP)and goose paramyxovirus disease,chimeric virus-like particles were con-structed and characterized based on the Newcastle disease virus-like particles(NDV VLPs)vector platform using the extracellular domain replacement strategy,insect baculovirus expression sys-tem,sucrose density gradient centrifugation and transmission electron microscopy.The VP3 gene of goose parvovirus(GPV)was linked to HN gene of Newcastle disease virus(NDV)and named rVP3,then the recombinant baculovirus rBV-rVP3 expressing rVP3 protein was constructed.After co-infection of sf9 cells with rBV-M and rBV-HN constructed previously in our laboratory for 72 h,the double chimeric virus-like particles GPV-NDV cVLPs were obtained.GPV-NDV cVLPs were purified by ultracentrifec-tion and sucrose gradient centrifugation,and the morphology of cV-LPs was similar to that of wild NDV under transmission electron microscope.Western blot tech-nique was.used to identify the proteins of each component of GPV-NDV cVLPs,and the results showed that the protein expression was correct.This study provides a safe and novel vaccine candi-date for the prevention of GP and goose paramyxovirus disease.
