摘要
为确定番鸭呼肠孤病毒(DRV)p10.8蛋白的生物学功能,本研究采用RT-PCR方法扩增DRVS14株p10.8编码基因,将其克隆到真核表达载体中,构建了重组表达质粒pcDNA-p10.8;通过转染鸭胚成纤维细胞(DEF),首次对p10.8蛋白的凋亡功能进行了研究。细胞转染48h后,Hoechest、DNALadder和TUNEL法的细胞凋亡检测结果显示:光镜下可见细胞形态学上出现的细胞皱缩,Hoechest染色后可见细胞核固缩,染色质凝固成团块状;DNALadder法可检测到凋亡细胞DNA样品呈梯形条带;TUNEL法可观察到褐色调亡细胞的存在。以上结果均表明,p10.8在DEF中的表达具有诱导宿主细胞凋亡的作用,是番DRV的凋亡蛋白。
The mechanisms by which Muscovy duck reoviruses （DRV） kill susceptible cells and ultimately produce diseases in Muscovy duck remain poorly understood. In this study, the pl0.8-encoding gene, the first ORF （ORFI） of DRV, was cloned in pcDNA3.1 （pcDNA-pl0.8） and expressed in primary duck embryonate fibroblast （DEF） cells. The p10.8 protein induced apoptosis was verified by DNA ladder formation, TUNEL and nuclear staining with Hoechest in pcDNA-pl0.8 transfected DEF. The results indicated that p10.8 is an apoptin of DVR.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2009年第10期766-769,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
973项目(2005CB522905)
