摘要
目前疫苗免疫是预防蓝舌病的主要有效手段,蓝舌病病毒样颗粒(VLPs)疫苗是蓝舌病疫苗研究的热点之一。为了制备蓝舌病病毒血清16型(BTV-16)病毒样颗粒,研究对载体pFastBac-Dual进行了改造,使其具有4个独立启动子的多克隆位点。依次将BTV-16 VP2、VP5、VP3和VP7基因插入到改造后载体(pF4)的4个多克隆位点中,成功构建了四表达重组质粒pF4-VP2-VP5-VP3-VP7,经转座获得重组杆粒Bacmid-VP2-VP5-VP3-VP7。通过转染Sf9细胞,获得可表达4种蛋白的重组杆状病毒rBac-VP2-VP5-VP3-VP7。经重组杆状病毒感染Sf9细胞,可共表达蛋白VP2、VP5、VP3和VP7,并通过蔗糖梯度离心制备出具有典型特征的BTV-16病毒样颗粒。
At present,vaccination is an effective method for the control of bluetongue(BT).As a new vaccine,bluetongue virus-like particles(BTV VLPs)has been a research hotspot.To synthesize BTV-16 VLPs,the vector pFastBac-dual was modified to have four multiple cloning sites(MCS)with independent promoters and named pF4.The VP2,VP5,VP3 and VP7 gene of BTV-16 were inserted into four MCSs of pF4,yielding the quadruple expression vectors pF4-VP2-VP5-VP3-VP7.After transposition,the recombinant bacmid Bacmid-VP2-VP5-VP3-VP7 was obtained.The recombinant insect baculovirus rBac-VP2-VP5-VP3-VP7 co-expressing four BTV-16 proteins(VP2,VP5,VP3 andVP7)was obtained by transfecting Sf9 cells.BTV-16 constructive proteins VP2,VP5,VP3 and VP7 were co-expressed and assembled into BTV-16 VLPs in infected Sf9 cells.BTV-16 VLPs were purified by sucrose density-gradient centrifugation and were observed to be empty and double-shelled under the electron microscope.
作者
黄超华
花群义
曹琛福
史卫军
阮周曦
林彦星
叶奕优
陶虹
王潇
陈金顶
HUANG Chaohua;HUA Qunyi;CAO Chenfu;SHI Weijun;RUAN Zhouxi;LIN Yanxing;YE Yiyou;TAO Hong;WANG Xiao;CHEN Jinding(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;Animal and Plant Inspection and Quarantine Technology Center,Shenzhen Customs District P.R.C.,Shenzhen,Guangdong 518045,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2021年第1期1-7,共7页
Chinese Journal of Veterinary Science
基金
国家“十三五”重点研发计划资助项目(2017YFD0502304,2017YFD0501104)。
