鹅细小病毒-鹅副黏病毒二联嵌合型病毒样颗粒的构建及鉴定  

Construction and identification of chimeric virus-like particles of goose parvovirus and goose paramyxovirus

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 SHAN Chunhui;LI Jindou;WU Jiaqi;FENG Jiaxuan;DING Jiaxin;CHEN Kainan;GUO Chunhong;DING Zhuang(Ministry of Education Key Laboratory for Zoonoses Research,College of Veterinary Medicine,Jilin University,Changchun 130062,China;College of Clinical Medicine,Changchun University of Traditional Chinese Medicine,Changchun 130117,China)

机构地区：[1]吉林大学动物医学学院人兽共患病研究教育部重点实验室,吉林长春130062 [2]长春中医药大学临床医学院,吉林长春130117

出　　处：《中国兽医学报》2023年第11期2237-2242,2307,共7页Chinese Journal of Veterinary Science

基　　金：长春市科技局重点研发计划资助项目(21ZGN17);国家自然科学基金资助项目(32072860);吉林省科技厅重点研发资助项目(20230202082NC)。

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 parvovirus,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 parvovirus 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。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 constructed and characterized based on the Newcastle disease virus-like particles(NDV VLPs)vector platform using the extracellular domain replacement strategy,insect baculovirus expression system,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 cVLPs was similar to that of wild NDV under transmission electron microscope.Western blot technique 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 candidate for the prevention of GP and goose paramyxovirus disease.

关 键 词：鹅细小病毒病 鹅副黏病毒病 VP3基因 嵌合型VLPs 

分 类 号：S852.65[农业科学—基础兽医学]