过表达非洲猪瘟病毒D1133L蛋白的MA-104细胞系建立及其初步应用  

Establishment and Preliminary Application of MA-104 Cell Line Overexpressing African Swine Fever Virus D1133L Protein

作　　者：张婷 杨博 崔卉梅 袁兴国 赵登率 杨金柯 郝雨 陈学辉 闫文倩 申超超 史喜绢 张大俊 杨行 刘湘涛[1] 郑海学[1] 张克山[1] ZHANG Ting;YANG Bo;CUI Huimei;YUAN Xingguo;ZHAO Dengshuai;YANG Jinke;HAO Yu;CHEN Xuehui;YAN Wenqian;SHEN Chaochao;SHI Xijuan;ZHANG Dajun;YANG Xing;LIU Xiangtao;ZHENG Haixue;ZHANG Keshan(State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China)

机构地区：[1]中国农业科学院兰州兽医研究所,兰州大学动物医学与生物安全学院家畜疫病病原生物学国家重点实验室,兰州730000

出　　处：《畜牧兽医学报》2022年第6期1877-1885,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划(2021YF1D1801300);甘肃省重大专项(20ZD7NA006,21ZD3NA001);中国农业科学院基本科研业务费(Y2020XK09);国家自然科学基金联合项目(31941002)。

摘　　要：旨在研究非洲猪瘟病毒(African swine fever,ASFV)编码的D1133L基因功能,本文利用慢病毒表达系统构建了过表达D1133L的MA-104/D1133L细胞系,分析ASFV在MA-104/D1133L细胞系与MA-104细胞中的复制差异。利用RT-qPCR和Western blot技术分别检测了ASFV感染MA-104/D1133L细胞后p30和p72基因的转录和蛋白表达水平,同时测定病毒滴度和病毒拷贝数评价ASFV的复制能力。结果表明:ASFV p30和p72基因在MA-104/D1133L细胞系的转录和蛋白表达水平高于野生型细胞系,ASFV在MA-104/D1133L细胞系的病毒增殖能力高于野生型细胞。本研究成功构建了过表达ASFV D1133L基因的MA-104/D1133L细胞系,与野生型细胞相比,D1133L能促进ASFV增殖,MA-104/D1133L细胞系更利于ASFV的复制,本结果为进一步研究ASFV D1133L基因功能积累了生物材料。To study the function of D1133L gene encoded by African swine fever virus(ASFV),we constructed MA-104/D1133L cell line for overexpression of D1133L using lentivirus expression vector,and compared ASFV replication ability in MA-104/D1133L and MA-104 cells.The transcription and protein expression levels of p30 and p72 in ASFV infected MA-104/D1133L cells were evaluated by real-time qPCR and Western blot respectively,and the proliferation of ASFV was evaluated according to virus titer and virus copies number.The transcriptional and protein levels of ASFV p30 and p72 proteins in MA-104/D1133L cell line were higher than those in wild MA-104 cell,and the viral proliferation capacity of ASFV in MA-104/D1133L cell line was higher than that in wild MA-104 cell line.The results showed that overexpressing ASFV D1133L gene of MA-104 cell line was successfully constructed in this study,that compared with wild type cells,MA-104/D1133L cell line was more conducive to ASFV replication,and D1133L promoted ASFV proliferation.These results provide biomaterials for further study of the function of ASFV D1133L gene.

关 键 词：非洲猪瘟病毒 D1133L基因 MA-104细胞 基因功能 过表达细胞系 

分 类 号：S852.659.6[农业科学—基础兽医学]