The study investigates the ability of subtype A Avian metapneumovirus (AMPV) to accept foreign genes and be used as a vector for delivery of Infectious bronchitis virus (IBV) QX genes to chickens. Initially the GFP gene was added to AMPV at all gene junctions in conjunction with the development of cassetted full length DNA AMPV copies. After recombinant virus had been recovered by reverse genetics, GFP positions supporting gene expression while maintaining virus viability in vitro, were determined. Subsequently, either S1 or nucleocapsid (N) genes of IBV were positioned between AMPV M and F genes, while later a bivalent recombinant was prepared by inserting S1 and N at AMPV MF and GL junctions respectively. Immunofluorescent antibody staining showed that all recombinants expressed the inserted IBV genes in vitro and furthermore, all recombinant viruses were found to be highly stable during serial passage. Eyedrop inoculation of chickens with some AMPV-IBV recombinants at one-day-old induced protection against virulent IBV QX challenge 3 weeks later, as assessed by greater motility of tracheal cilia from chickens receiving the recombinants. Nonetheless evidence of AMPV/IBV seroconversion, or major recombinant tracheal replication, were largely absent.

译文

:这项研究调查了A型禽亚肺炎病毒(AMPV)接受外源基因并用作将传染性支气管炎病毒(IBV)QX基因传递给鸡的载体的能力。最初,随着盒装全长DNA AMPV拷贝的发展,将GFP基因添加到AMPV的所有基因连接处。在通过逆向遗传学回收重组病毒之后,确定了在体外维持病毒生存力的同时支持基因表达的GFP位置。随后,IBV的S1或核衣壳(N)基因位于AMPV M和F基因之间,随后通过在AMPV MF和GL连接处分别插入S1和N制备二价重组体。免疫荧光抗体染色显示,所有重组体均在体外表达了插入的IBV基因,此外,还发现所有重组体病毒在连续传代过程中均高度稳定。一天后,用一些AMPV-IBV重组体对鸡进行眼药水接种可在3周后诱导出针对强毒IBV QX攻击的保护作用,这是通过接受重组体的鸡的气管纤毛运动性增强来评估的。但是,基本上没有AMPV / IBV血清转换或主要的重组气管复制的证据。

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