鸭肠炎病毒核衣壳蛋白质互作蛋白质基因荧光定量PCR检测方法的建立

为进一步阐明鸭肠炎病毒(DEV)的致病机理,本试验建立检测鸭肠炎病毒核衣壳蛋白质(NP)的互作蛋白质(PKCI)基因荧光定量PCR方法。针对PKCI基因设计特异性引物,经PCR扩增目的基因构建重组质粒p MD18-T-PKCI,将其作为阳性标准品构建荧光定量PCR标准曲线,并对建立的荧光定量PCR方法进行重复性、特异性和敏感性试验。结果显示:标准曲线的线性关系为Y=-3.31x+42.00,相关系数为-1.00,扩增效率为100%,熔解曲线仅出现单特异峰,对H5亚型、H7亚型和H9亚型禽流感病毒及新城疫病毒、鸭肝炎病毒均未检测到荧光信号。表明本试验建立的方法具有良好的稳定性、特异性和灵敏性。

猪圆环病毒3型检测及其Cap结构序列和抗原性预测分析

近期报道在美国多个州的养猪场发现了一种新型的猪圆环病毒3型毒株(porcine circovirus 3,PCV3)。为了确定PCV3是否已经在我国南方部分省份的猪场中广泛存在,并对目前PCV2疫苗是否对PCV3感染具有交叉保护作用作出科学预测,笔者对我国南方几个省份疑似猪皮炎肾病综合征的病猪组织样品进行PCV3巢式PCR检测及其核衣壳蛋白质(capsid protein,Cap)结构的PyMol和抗原表位服务器在线模拟分析。结果首次在这些病料中检测到PCV3的存在,部分病料表现为PCV2和PCV3共感染的特性。对其Cap结构及抗原性分析发现,PCV2和PCV3表现出很大的差异。因此,推测PCV2和PCV3不具有交叉免疫保护的特性,新的疫苗研究和开发,对于将来控制PCV3流行是必要的。

Host Cell Protein C9orf9 Promotes Viral Proliferation via Interaction with HSV-1 UL25 Protein

In light of the scarcity of reports on the interaction between HSV-1 nucleocapsid protein UL25 and its host cell proteins,the purpose of this study is to use yeast two-hybrid screening to search for cellular proteins that can interact with the UL25 protein.C9orf69,a protein of unknown function was identified.The interaction between the two proteins under physiological conditions was also confirmed by biological experiments including co-localization by fluorescence and immunoprecipitation.A preliminary study of the function of C9orf69 showed that it promotes viral proliferation.Further studies showed that C9orf69 did not influence viral multiplication efficiency by transcriptional regulation of viral genes,but indirectly promoted proliferation via interaction with UL25.