猫泛白细胞减少症病毒的分离鉴定及其结构蛋白VP2可溶性表达的调控  

Isolation and identification of feline panleukopenia virus and regulation of soluble expression of its structural protein VP2

作　　者：刘丽春 徐国伟 茹毅 李亚军 杨明星 梁超 刘晓飞 张世栋[2] 李建喜[2] 王学智[2] 孙研 周雨霞[1] LIU Li-chun;XU Guo-wei;RU Yi;LI Ya-jun;YANG Ming-xing;LIANG Chao;LIU Xiao-fei;ZHANG Shi-dong;LI Jian-xi;WANG Xue-zhi;SUN Yan;ZHOU Yu-xia(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences,Lanzhou 730050,China;Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050 [3]中国农业科学院兰州兽医研究所,甘肃兰州730046

出　　处：《中国兽医科学》2023年第6期706-714,共9页Chinese Veterinary Science

基　　金：甘肃省青年科技基金项目(21JR7RA035);兰州市人才创新创业项目(2022-RC-46);中国农业科学院基本科研业务费项目(Y2022XK19,1610032021017)。

摘　　要：为了研究猫泛白细胞减少症病毒(FPV)的生物学特性,可溶性表达FPV结构蛋白,研发基于VP2的新型亚单位疫苗,采集40份疑似FPV阳性的中华田园猫粪便样本,通过qPCR对样本检测后利用细胞培养、血凝试验、PCR和电镜观察等试验鉴定分离株,并对其VP2基因进行同源性比对。同时,构建pET-28a-VP2、pET-28a-SUMO-VP2和pET-22a-SUMO-VP2重组质粒并利用大肠杆菌进行表达。结果显示,成功分离出1株FPV毒株;生物信息学分析表明,该毒株的VP2基因与此前报道的毒株同源性高达99%。蛋白表达结果显示,单独使用SUMO标签对VP2蛋白的可溶性表达影响较低;然而,当带有SUMO标签的重组质粒与4个分子伴侣(pG-KJE8、pGro7、pKJE7、pTf16)分别共表达时,可溶性VP2的表达量可显著提高。该研究可为FPV亚单位疫苗研发提供数据和有效的生物材料。In order to develop a novel subunit vaccine based on feline panleukopenia virus(FPV)VP2,study the biological characteristics of FPV and solubly express VP2 structural proteins,forty faecal samples of Felis silvestris catus suspected to be FPV positive were collected,and the samples were detected by qPCR.The isolates were identified by cell culture,hemagglutination test,PCR and electron microscopy observation and the homology of VP2 gene was compared.Meanwhile,the recombinant plasmids of pET-28a-VP2,pET-28a-SUMO-VP2 and pET-22a-SUMO-VP2 were constructed and expressed by Escherichia coli.The results showed that an FPV strain was successfully isolated.Bioinformatics analysis showed that the VP2 gene of this strain had 99%homology with the stains previously.And the results showed that the soluble expression of VP2 protein was less affected by the use of SUMO tag alone.However,when the recombinant plasmid with SUMO tag was coexpressed with four molecular chaperones(pG-KJE8,pGro7,pKJE7,pTf16),the expression of soluble VP2 was significantly increased.This study can provide data and effective biomaterials for the development of FPV subunit vaccine.

关 键 词：猫泛白细胞减少症病毒 中华田园猫 SUMO标签 分子伴侣 亚单位疫苗 

分 类 号：S852.659.2[农业科学—基础兽医学]