非洲猪瘟病毒MGF300-1L蛋白的生物学特性分析  

Biological characterization of MGF300-1L protein of Africanswine fever virus

作　　者：罗瑞 王涛[2] 钟代浪 潘力[2] 孙茂文 孙元[2] 黄淑坚[1] 仇华吉 LUO Rui;WANG Tao;ZHONG Dailang;PAN Li;SUN Maowen;SUN Yuan;HUANG Shujian;QIU Huaji(College of Life Science and Engineering,Foshan Institute of Science and Technology,Foshan,Guangdong 528231,China;Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)

机构地区：[1]佛山科学技术学院、生命科学与工程学院,广东佛山528231 [2]中国农业科学院、哈尔滨兽医研究所、兽医生物技术国家重点实验室,黑龙江哈尔滨150069

出　　处：《中国兽医学报》2023年第1期1-9,共9页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金面上资助项目(32072854)。

摘　　要：为了解非洲猪瘟病毒(African swine fever virus,ASFV)编码的未知功能蛋白MGF300-1L的基本生物学特性,利用生物信息学手段分析了该蛋白的保守性、理化性质,并对其翻译后修饰、结构与功能进行预测;通过实时荧光定量PCR分析在病毒感染条件下MGF300-1L的表达特性;最后,利用MGF300-1L的真核表达质粒,通过激光共聚焦试验分析其亚细胞定位,并采用间接免疫荧光试验(IFA)分析其免疫原性。结果显示,MGF300-1L蛋白在基因Ⅱ型ASFV中高度保守,为碱性疏水膜蛋白,具有糖基化和磷酸化修饰,与G蛋白偶联受体激酶相互作用蛋白2(GIT2)结构的同源性为21%;MGF300-1L为ASFV的早期基因,其编码蛋白主要定位于细胞质;IFA未检测到ASFV感染猪的阳性血清中存在针对MGF300-1L蛋白的抗体。本研究发现,MGF300-1L为ASFV高度保守的非必需早期基因,其编码蛋白具有跨膜区及糖基化、磷酸化修饰,并定位于细胞质,免疫原性较差,为进一步探究MGF300-1L蛋白在ASFV复制周期中的功能奠定了基础。To understand the basic biological characteristics of the unidentified protein MGF300-1L encoded by the MGF300-1L gene of African swine fever virus(ASFV),the conservation and physicochemical properties of the MGF300-1L protein was first analyzed by bioinformatics methods.The posttranslational modification,structure and function of the MGF300-1L protein were also predicted.Then,the expression characteristics of the MGF300-1L under viral infection were analyzed by real-time PCR.Finally,the subcellular localization of the MGF300-1L protein was analyzed by laser confocal assay,and the immunogenicity was investigated by indirect immunofluorescence assay(IFA).The results showed that the MGF300-1L protein was highly conserved in gene typeⅡASFV.It was an alkaline hydrophobic membrane protein with glycosylation and phosphorylation modification sites,and the homology of G protein-coupled receptor kinase interaction protein(GIT2)structure was 21%.MGF300-1Lwas an early expressed gene of ASFV,and its encoding protein mainly located in the cytoplasm.The antibodies against MGF300-1L protein in positive serum were not detected by IFA.In this study,it was found that MGF300-1L was a highly conserved unnecessary early expressed gene of ASFV,and its encoding protein had transmembrane region,glycosylation and phosphorylation modification,and located in the cytoplasm,the immunogenicity was poor.These results lay the foundation for further exploring the function of the MGF300-1L protein in the ASFV replication cycle.

关 键 词：非洲猪瘟病毒 MGF300-1L蛋白 亚细胞定位 免疫原性 

分 类 号：S852.65[农业科学—基础兽医学]