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首页> 外文期刊>Revista Argentina de Microbiologia >Expression of the hemagglutinin HA1 subunit of the equine influenza virus using a baculovirus expression system
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Expression of the hemagglutinin HA1 subunit of the equine influenza virus using a baculovirus expression system

机译:使用杆状病毒表达系统表达马流感病毒血凝素HA1亚基

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摘要

Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens’ eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for producing recombinant equine influenza hemagglutinin to be used as a vaccine. The hemagglutinin ectodomain (HA1 subunit) was cloned and expressed using a baculovirus expression vector. The expression was determined by SDS-PAGE and immunoblotting. A high yield, 20μg/ml of viral protein, was obtained from recombinant baculovirus-infected cells. The immune response in BALB/c mice was examined following rHA1 inoculation. Preliminary results show that recombinant hemagglutinin expressed from baculovirus elicits a strong antibody response in mice; therefore it could be used as an antigen for subunit vaccines and diagnostic tests.
机译:马流感病毒是全球马匹呼吸道疾病的主要原因。通过灭活全病毒疫苗接种来预防疾病。目前,大多数流感疫苗都是在有胚鸡蛋中产生的。从尿囊液中收获病毒颗粒并进行化学灭活。尽管该系统已经使用了很多年,但使用鸡蛋作为疫苗生产的基质却有几个公认的缺点(成本,鸡蛋供应,废物处置和鸡蛋产量)。这项研究的目的是评估杆状病毒系统,作为生产重组马流感血凝素用作疫苗的潜在方法。使用杆状病毒表达载体克隆并表达血凝素胞外域(HA1亚基)。通过SDS-PAGE和免疫印迹确定表达。从重组杆状病毒感染的细胞中获得了高产量的20μg/ ml病毒蛋白。接种rHA1后检查了BALB / c小鼠的免疫反应。初步结果表明,杆状病毒表达的重组血凝素在小鼠中引起强烈的抗体反应。因此,它可用作亚单位疫苗和诊断测试的抗原。

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