过表达HDAC6基因的Vero细胞系建立及其对狂犬病病毒增殖效率评价  

Establishment of Vero cell lines overexpressing HDAC6 gene and evaluation of its proliferative efficiency of rabies virus

作　　者：殷娟斌 张志雄 王莎莎 马小琴 梁正基 孙跃峰 王相伟 殷相平[1] YIN Juan-bin;ZHANG Zhi-Xiong;WANG Sha-sha;MA Xiao-qin;LIANG Zheng-ji;SUN Yue-feng;WANG Xiang-wei;YIN Xiang-ping(State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2023年第2期150-155,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(32202779);甘肃省自然科学基金项目(21JR7RA022)。

摘　　要：为提高狂犬病病毒(rabies virus,RABV)在细胞中的增殖效率,本研究利用慢病毒包装系统构建组蛋白去乙酰化酶6(histone deacetylase 6,HDAC6)过表达的Vero/HDAC6细胞系,分析RABV在Vero/HDAC6细胞系与Vero细胞中的复制差异。首先以非洲绿猴HDAC6基因为目标构建重组质粒plv-SV40-puro-3×flag-HDAC6,并与辅助质粒pMD2.G和psPAX2共转染至HEK-293T细胞进行慢病毒包装。将包装好的慢病毒感染Vero细胞并通过嘌呤霉素筛选获得过表达HDAC6基因的阳性细胞。利用TCID50和Western-blot技术检测RABV在Vero/HDAC6细胞系中的复制能力。结果表明,过表达HDAC6能够明显促进RABV复制,并且RABV在Vero/HDAC6细胞系中的增殖能力高于野生型细胞。本研究构建的Vero/HDAC6细胞系能显著促进RABV的增殖,为RABV疫苗候选细胞株奠定了基础。In order to improve the proliferation efficiency of Rabies virus(RABV)in cells,a Vero/HDAC6cell line overexpressing histone deacetylase 6(HDAC6)was constructed using lentivirus packaging system in this study and compared RABV replication ability in Vero/HDAC6 and Vero cells.The recombinant plasmid plv-SV40-puro-3×flag-HDAC6 was constructed with African green monkey HDAC6 gene as the target,and co-transfected with helper plasmids p MD2.G and ps PAX2 into HEK-293T cells for lentiviral packaging.Vero cells were infected with the packaged lentivirus and the HDAC6 overexpressing cells were screened by puromycin antibiotic.The replication efficiency of RABV in Vero/HDAC6 cell line was detected by TCID50 and Western-blot.The results showed that overexpression of HDAC6 can promote the replication of RABV,and the proliferation ability of RABV in Vero/HDAC6 cell line was higher than that in wild-type cells.The Vero/HDAC6 cell line constructed in this study can significantly promote the proliferation of RABV,which provides a candidate cell line for RABV vaccine productions.

关 键 词：狂犬病病毒 HDAC6 VERO细胞 慢病毒包装 过表达细胞系 

分 类 号：S859.659.5[农业科学—临床兽医学]