本文選題：新城疫病毒 + 基因Ⅶ型　； 參考：《中國獸醫(yī)學報》2017年02期

【摘要】：利用昆蟲細胞/桿狀病毒表達載體系統(tǒng),將已構建編碼新城疫病毒(NDV)M、F和HN結構基因的3種單順反子重組桿粒共感染懸浮培養(yǎng)的Sf9細胞以大量制備新城疫病毒樣顆粒(ND VLPs)。收集細胞培養(yǎng)上清經(jīng)超速離心濃縮,蔗糖密度梯度純化后,利用Western blot檢測和透射電鏡觀察。結果顯示,3種目的蛋白均正確表達且組裝形成與原病毒粒子形態(tài)相似的顆粒,測定其血凝活性效價可達12log2。動物試驗結果表明,與LaSota滅活油乳苗相比,將純化后的病毒樣顆粒以含25μg蛋白的劑量或聯(lián)合佐劑免疫商品蛋雞后能產(chǎn)生較高水平的HI抗體,且可有效減少攻毒雞只口咽、泄殖腔途徑的排毒時間,從而減少病毒在健康雞群中的傳播感染機會;表明所研制的基因Ⅶ型ND VLPs可作為疫苗候選株具有很大的防控優(yōu)勢。
[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，

本文編號：1780382