脑心肌炎病毒受体的筛选、鉴定及功能分析

脑心肌炎病毒是一种重要人畜共患病病原体，自然宿主和分布地域都比较广泛，该病毒主要引起人畜的急性心肌炎、糖尿病等，并可导致母畜流产、仔畜死亡，严重危害人类健康与畜牧业生产。该病毒受体类型、病毒与宿主细胞之间相互作用机制至今仍不明确。本项目拟采用酵母双杂交、病毒覆盖蛋白印记和质谱分析技术系统地筛选脑心肌炎病毒候选受体；运用敏感细胞受体功能丧失、非敏感细胞受体功能获得对候选受体进行鉴定；运用基因分段截短、突变结合GST Pull-down技术研究病毒的结合位点（区域），并分析病毒和宿主细胞作用关系。以期能阐明该病毒的受体类型和病毒与宿主细胞之间相互作用机制，为进一步研究脑心肌炎病毒的感染与免疫机制、致病性以及防治技术奠定基础。

Encephalomyocarditis virus (EMCV) is an important zoonotic virus, which has a wide range of natural hosts and distribution areas. EMCV causes acute myocarditis,diabetes both in human and animals,as well as the abortion of pregnant female animals and the newborn death, hazarding on human health and animal husbandry potentially. In recent years, there is few research about the EMCV receptors,even their types and the interaction mechanism between the virus and the receptors are also not clear. The present study is to employ yeast two-hybrid technology,viral overlay protein blot assay technology, mass spectrometry to screen the virus receptors, through sensitive cell defunctionalization, nonsensitive cell function acquisition and RT-PCR techniques to verify the EMCV receptors, and study the binding site by using gene fragmentation, site-specific mutagenesis and GST Pull-down technology, so as to clarify the interaction mechanism between the virus and host cells, and lay the foundation for the research of virus infection and the immune mechanism,pathogenicity and the related prevention technology.