血红蛋白β亚基对猪流行性腹泻病毒增殖抑制作用的研究  

Inhibition of porcine epidemic diarrhea virus replication by hemoglobin β-subunit in Vero cells

作　　者：徐云飞[1,2] 郭龙军[2] 张健[2] 时洪艳[2] 冯力[2] 任慧英[1] 王玉娥[2] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [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150001

出　　处：《中国预防兽医学报》2016年第8期605-608,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金面上项目(31572497);黑龙江省留学归国科学基金项目(LC201418)

摘　　要：本实验室前期蛋白质组学研究结果显示,猪流行性腹泻病毒(PEDV)感染非洲绿猴肾细胞(Vero-E6)后,导致血红蛋白β亚基(HBB)的表达量升高。为进一步从蛋白水平验证前期的实验结果并研究HBB对PEDV增殖作用的影响,本研究将PEDV感染Vero-E6细胞,感染24 h后经western blot检测结果表明,PEDV感染能够促进HBB的表达。进一步采用RT-PCR从Vero-E6细胞总RNA中扩增HBB基因并将其克隆至真核表达载体pCAGGS中,构建HBB真核表达重组质粒pCAGGS-HBB,并将其转染至Vero-E6细胞。经western blot检测结果表明,HBB在Vero-E6细胞中获得高效表达。在HBB过表达后感染PEDV,经western blot、TCID_(50)及荧光定量PCR检测结果表明,过表达HBB能够显著抑制PEDV的增殖。本研究对于进一步阐明HBB对PEDV增殖作用的影响机制提供了实验依据。Our previous study showed that porcine epidemic diarrhea virus （PEDV） infection upregulates the expression of hemoglobin β-subunit （HBB） in Vero-E6. To further study the association between HBB and PEDV infection, the HBB gene was amplified by RT-PCR from Vero-E6 and cloned into the eukaryotic expressing plasmid of pCAGGS. Then the resultant recombinant plasmid was transfected into Vero-E6 cells for overexpression, and the cells were infected with PEDV at 24 hrs post the transfection. By using western blot, real time RT-PCR and TCID50 assays, the results shown that the overexpression of HBB was able to significant inhibit PEDV replication in the cells. In conclusion, our finding laid a foundation to further clarify the impact of HBB on PEDV proliferation.

关 键 词：血红蛋白β亚基 猪流行性腹泻病毒 病毒增殖 

分 类 号：S852.65[农业科学—基础兽医学]