O型口蹄疫病毒P1-2A3C基因在家蚕杆状病毒表达系统中的表达

口蹄疫是一种严重危害畜牧业生产的烈性传染病。为了促进O型口蹄疫病毒(FMDV)基因工程活载体疫苗的研制,选取O型FMDV编码序列中的衣壳蛋白前体P1-2A基因和蛋白酶3C基因,插入家蚕杆状病毒转移载体pVL1393中,构建重组载体pVL-P1-2A3C,并与线性化病毒Bm-BacPAK6 DNA共转染家蚕BmN细胞,获得重组病毒Bm-P1-2A3C。将重组病毒感染家蚕5龄幼虫,以双抗体夹心ELISA法和间接血凝方法检测血淋巴中的表达产物:目的蛋白在感染病毒后120 h的蚕血淋巴中表达量最高,抗原表达呈阳性的最大稀释倍数为1∶128。结果显示O型FMDV的P1-2A3C基因已在家蚕体内获得表达。

Expression of FMDV Type O p1-2A3C Gene in Bombyx mori Baculovirus Expression Vector System

Food and mouth disease is one of fatal infectious diseases which jeopardizes animal husbandry seriously.In order to develop the genetic engineering live vector vaccine of type O FMDV,procusor P1-2A and protease 3C was selected from the encoding sequences of type O,which was inserted into Bombyx mori baculovirus transfer vector pVL1393 and the recombinant vector pVL-P1-2A3C was constructed.The recombinant vector was co-transfected with linear DNA of Bm-BacPAK6 virus into BmN cells.Then the recombinant virus Bm-P1-2A3C was gained.Bm-P1-2A3C infected the silkworm larvae at 5th instars,and the expressed product in haemolymph was determined by double-antibody sandwich ELISA and indirect hemagglutination test.The results showed that the highest detectable level of target protein in haemolymph was at 120 h post infection,and the maximum dilution was 1∶128 with positive expression of antigen.The above results indicated that P1-2A3C gene of type O FMDV have been obtained in Bombyx mori baculovirus expression system.