牛疱疹病毒1型复制非必需基因UL0.5的鉴定  

The UL0.5 gene is a non-essential gene for bovine herpesvirus-1 replication

作　　者：尹贻鑫[1] 何宁宇[1] 张瑞竹[1] 张炜[1] 陈兴江[1] 陈焕春[1] 刘正飞[1] 

机构地区：[1]华中农业大学动物医学院农业微生物国家重点实验室,湖北武汉430070

出　　处：《中国预防兽医学报》2013年第11期865-869,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31070133)

摘　　要：牛疱疹病毒1型(BHV-1)和5型(BHV-5)为同源性最高的两种α疱疹病毒亚科成员,分别引起牛的呼吸道和神经症状。序列比较显示,BHV-1基因组中具有一个独特的UL0.5基因。为研究该基因的功能,本研究构建BHV-1 UL0.5基因缺失转移质粒,与野生型基因组共转染MDBK细胞,通过空斑纯化获得UL0.5基因缺失突变株(rBHV-1ΔUL0.5);PCR和western blot鉴定结果表明,重组病毒形成的空斑大小及一步法生长曲线与野生型基本相同;家兔接种试验表明,突变株与野生型在急性感染期、潜伏感染期及再激活期病毒排毒量无明显变化;Real time PCR检测3个时期三叉神经节中病毒基因组的拷贝数也无明显差别。综上所述,UL0.5基因是BHV-1的复制非必需基因,该基因缺失后对家兔致病性无显著影响。Bovine herpesvirus 1 （BHV-1） and bovine herpesvirus 5 （BHV-5） are very similar in viral genomic sequence, but they cause different symptoms in infected cattle, upper respiratory tract disorder for BHV-1 and nervous symptoms for BHV-5. Comparing the two viral genomic sequences, we found that an UL0.5 gene is only present in BHV-1 genome, but not in BHV-5 genome. To study the biological characterization of the UL0.5 gene, we constructed a transfer plasmid to eliminate the UL0.5 gene and co-transfected with wild-type genomic DNA of BHV-1 into MDBK cells to generated the UL0.5 gene deleted BHV-1 （rBHV-1AUL0.5）, which was identified by PCR and western blot. Comparison of plaque size and one-step growth curve showed that the proliferation titer of the mutant and the wild type virus were in the same level. In a^tdition, the viral shedding changes of the mutant and the wild type virus were not significant in the phases of acute infection, latency, and reactivation in rabbits artificially infected with the two virus, respectively. Furthermore, there was also no significant difference in the viral copy numbers in the trigeminal ganglia of the three phases detected by real time PCR assay, and the rBHV-1AUL0.5 had little impact to the pathogenicity in rabbit. In conclusion, the UL0.5 gene is a non-essential gene for BHV-1 proliferation.

关 键 词：牛疱疹病毒1型 UL0.5基因 增殖特性 致病性 

分 类 号：S852.65[农业科学—基础兽医学]