基因Ⅶ型新城疫病毒样颗粒作为疫苗候选株的免疫效果

发布时间：2018-04-21 01:36

  本文选题：新城疫病毒 + 基因Ⅶ型　； 参考：《中国兽医学报》2017年02期

【摘要】：利用昆虫细胞/杆状病毒表达载体系统,将已构建编码新城疫病毒(NDV)M、F和HN结构基因的3种单顺反子重组杆粒共感染悬浮培养的Sf9细胞以大量制备新城疫病毒样颗粒(ND VLPs)。收集细胞培养上清经超速离心浓缩,蔗糖密度梯度纯化后,利用Western blot检测和透射电镜观察。结果显示,3种目的蛋白均正确表达且组装形成与原病毒粒子形态相似的颗粒,测定其血凝活性效价可达12log2。动物试验结果表明,与LaSota灭活油乳苗相比,将纯化后的病毒样颗粒以含25μg蛋白的剂量或联合佐剂免疫商品蛋鸡后能产生较高水平的HI抗体,且可有效减少攻毒鸡只口咽、泄殖腔途径的排毒时间,从而减少病毒在健康鸡群中的传播感染机会;表明所研制的基因Ⅶ型ND VLPs可作为疫苗候选株具有很大的防控优势。
[Abstract]:By using insect cell / baculovirus expression vector system, three kinds of eukaryotic co-infected Sf9 cells encoding NDV MU F and HN structural genes of Newcastle disease virus (NDV) were constructed to produce Newcastle disease virus (NDV) like particles. The supernatant of cell culture was concentrated by ultracentrifugation and purified by sucrose density gradient. The supernatant was detected by Western blot and observed by transmission electron microscope (TEM). The results showed that all three target proteins were correctly expressed and assembled to form particles similar to those of protovirus particles. The hemagglutination activity titer of the three proteins was up to 12 log2. The results of animal experiment showed that compared with LaSota inactivated oil emulsion vaccine, the purified virus-like particles could produce a higher level of HI antibody after immunizing layer hens with 25 渭 g protein or adjuvant, and could effectively reduce the oral pharynx of infected chickens. The detoxification time of cloacal pathway reduces the chance of transmission and infection of the virus in healthy chickens, which indicates that the ND VLPs gene 鈪，

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