马疱疹病毒1型gD基因主要中和抗原区在大肠埃希氏菌中的表达

利用PCR技术扩增马疱疹病毒1型(EHV-1)gD基因主要中和抗原编码区,将其克隆到原核表达载体pGEX-6p-1中,获得的重组质粒pGEX-EHV-gD用IPTG诱导表达。SDS-PAGE电泳分析显示,重组gD蛋白以不溶性包涵体形式表达,与预期大小(43 ku)相符;Western-blotting分析表明,表达产物均能与抗EHV-1和EHV-4阳性血清发生反应。用重组蛋白免疫家兔获得的超免疫血清能中和EHV-1和EHV-4,且中和抗体效价较高,说明原核表达产物具有良好的免疫原性,可以用作ELISA包被抗原或重组基因疫苗。     

马动脉炎病毒RT-LAMP检测方法的建立

为建立一种快速、简便、特异性高的马动脉炎病毒反转录环介导等温扩增(RT-LAMP)检测方法,针对马动脉炎病毒膜基质蛋白M基因的保守序列设计特异性引物,优化反应条件,建立了马动脉炎病毒的RT-LAMP检测方法并对其特异性和灵敏性进行了检测。结果显示,建立的RT-LAMP检测方法具有良好的特异性,灵敏度为0.12pg,是普通RT-PCR的100倍,检测时间仅需1h,肉眼即可观察检测结果。表明建立的RT-LAMP检测方法可用于进出口马动脉炎病毒的检疫。     

马疱疹病毒糖蛋白G基因C-末端的原核表达

为获得马疱疹病毒1型(EHV1)和4型(EHV4)糖蛋白G(gG),对克隆载体pMD-1gG和pMD-4gG分别进行BamHⅠ+SalⅠ和EcoRⅠ+HindⅢ双酶切,将所得片段分别插入表达载体pGEX-6P-1和pET28a(+)中,构建了重组质粒pGEX-1gG和pET-4gG。经双酶切和序列鉴定为阳性后,将重组质粒pGEX-1gG转化入大肠杆菌BL21(DE3)、重组质粒pET-4gG转化入大肠杆菌Rosetta(DE3)中,经IPTG诱导表达后对表达产物进行SDS-PAGE分析。结果显示,分别在35和17ku处出现与预期大小相符的目的条带。Western-blot结果证实,目的蛋白均具有良好的反应原性。表明,成功实现了EHV1gG和EHV4gG蛋白的正确表达,初步鉴定其具有良好的反应原性。     

Horsing around: Using equine facilitated learning to support the development of social-emotional competence of students at risk of school failure

The social-emotional issues some students experience can place them at risk of school failure. Traditional methods of support can be ineffective or not sustainable and new alternative approaches need to be attempted to support social-emotional competency, school engagement, and success for students at risk. This article discusses preliminary outcomes of an equine facilitated learning (EFL) program specifically designed to focus on using horses to improve the resilience and social-emotional competency in students perceived as “at risk” of school failure. This qualitative exploratory study used interviews and observations over a six month period to listen to the voices of the students themselves about their experiences of EFL. Initial findings from the pilot study suggest that EFL programs can be a novel and motivating way to promote resilience training and social-emotional development of students at risk of failure and, in turn, improve their level of engagement and connection with school environments.     

马鼻肺炎疫苗的研究现状及前景分析

介绍了3类马鼻肺炎疫苗的制备工艺、自身特点,分析比较了各类型疫苗的优缺点,并对其商品化进行了可行性分析,指出传统疫苗(灭活疫苗、减毒疫苗)是目前传染病防治的主要产品。虽然新型疫苗(病毒活载体疫苗)还处在研究阶段,但终将成为马鼻肺炎疫苗未来发展的主要方向。     

应用多重PCR检测和区分3个型的马疱疹病毒

针对马疱疹病毒(EHV)的EHV-1、EHV-2和EHV-4糖蛋白B基因序列,设计、合成了3对特异性引物进行多重PCR,不仅可以在数小时内分别检测这3个型的EHV,而且在同一反应系统内可以清晰地区分EHV-1、EHV-2和EHV-4,其PCR产物大小分别为226、3335、70bp,符合预期的片段大小,序列分析证实与已发表的序列一致;该检测方法的灵敏度达到103TCID50;分别从血清学阳性但病毒分离为阴性的1匹进口马组织样品和一些出口前检疫马的鼻咽样品检测到EHV-1和EHV-4特异性核酸。