传染性法氏囊病病毒的病毒样颗粒制备及免疫原性分析  

Preparation and immunogenicity analysis of virus-like particles of infectious bursal disease virus

作　　者：黄建飞 申翰钦 张钰坤 巫秀红 黄松健 严专强 周庆丰 HUANG Jianfei;SHEN Hanqin;ZHANG Yukun;WU Xiuhong;HUANG Songjian;YAN Zhuanqiang;ZHOU Qingfeng(Wen's Foodstuff Group Co.,Ltd.,Yunfu 527400,China;Key Laboratory of Healthy Breeding of Livestock and Poultry and Environmental Control,Guangdong Province Department of Science and Technology,Yunfu 527400,China;College of Animal Sciences,South China Agricultural University,Guangzhou 510642,China;College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]温氏食品集团股份有限公司,广东云浮527400 [2]广东省畜禽健康养殖与环境控制企业重点实验室,广东云浮527400 [3]华南农业大学动物科学学院,广东广州510642 [4]华南农业大学兽医学院,广东广州510642

出　　处：《畜牧与兽医》2021年第6期67-72,共6页Animal Husbandry & Veterinary Medicine

摘　　要：为了研究传染性法氏囊病病毒(IBDV)病毒样颗粒(VLP)的免疫原性,通过RT-PCR扩增其VP2基因,克隆到pSYno-1原核表达载体上,转化到大肠杆菌BL21(DE3)感受态细胞进行表达。通过烟草蚀纹病毒(tobacco etch virus,TEV)蛋白酶酶切后制备VLP并进行免疫原性分析。结果显示:His-MBP标签能够促进VP2蛋白的可溶性表达,其可溶性表达水平约为50%;透射电镜观察发现TEV蛋白酶切开的VP2能形成25 nm左右的VLP;动物试验结果发现VLP免疫能诱导产生较高的抗体水平,并能保护免疫鸡抵抗IBDV强毒攻击,其中免疫组脾体比显著低于非免疫组(P<0.05),免疫组血清和组织中病毒含量极显著低于非免疫组(P<0.01),表明VLP具有良好的免疫原性。本研究为IBDV基因工程疫苗的研发提供了参考。In order to identify the immunogenicity of the virus-like particles(VLP)of the infectious bursal disease virus(IBDV),the VP2 gene of IBDV was amplified by RT-PCR and cloned onto a pSYNO-1 prokaryotic expression vector.The recombinant vector was transformed into E.coli BL21(DE3)competent cells for recombinant VP2 protein expression.The recombinant VP2 protein was digested by TEV protease to form VLP.Finally,the immunogenicity of the VLP was evaluated in SPF chickens.The results showed that the His-MBP tag promoted the solubility of the VP2 protein,and the soluble protein level was about 50%.Transmission electron microscopy(TEM)observation showed that the VP2 digested by TEV protease formed VLP of about 25 nm.The animal experiment showed that immunization with the VP2-based VLP induced a high level of specific antibodies and protected SPF chickens against the challenge by the virulent IBDV.The spleen body ratio of the immunized group of chickens was significantly lower than that of the non-immunized group(P<0.05),and the viral content in the serum and tissues of the immunized group was significantly lower than that of the non-immunized group(P<0.01),which indicated excellent immunogenicity of VLP.This study laid a foundation for development of vaccines of IBDV in genetic engineering.

关 键 词：传染性法氏囊病毒 VP2蛋白 融合标签 原核表达 病毒样颗粒 免疫原性 

分 类 号：S852.6[农业科学—基础兽医学]