EXPRESSION OF ROTAVIRUS OUTER CAPSID PROTEIN VP4 BY RECOMBINANT ADENOVIRUS

猪轮状病毒OSU外衣壳蛋白VP4被选为 研究该蛋白免疫原性的模型 腺病毒载体。腺病毒经口给药的能力 使腺病毒载体有可能用于疫苗生产 特别吸引人。此外，活体腺病毒疫苗已经 已经证明口服给药对人类是安全的 肠溶胶囊。 克隆了猪轮状病毒OSU株VP4全长基因. 在E3下游克隆入腺病毒5型 推动者。在这个活的重组体中，轮状病毒VP4序列是 取代了腺病毒E3区的序列。这块牌匾- 纯化的重组(Ad5-OSU VP4)，表达了一种抗原性可靠的 免疫沉淀法测定VP4蛋白。84 kDa蛋白质 具有与OSU轮状病毒天然VP4相同的凝胶迁移率 在MA104细胞中生长。一项研究的初步结果，其中棉花 用Ad5-OSU VP4活体鼻腔感染大鼠，结果表明 动物产生低水平的OSU中和抗体 猪轮状病毒。

The outer capsid protein VP4 of porcine rotavirus OSU was chosen as a model to study the immunogenicity of this protein expressed by an adenovirus vector. The ability of adenovirus to be administered orally makes the potential use of adenovirus vectors for vaccine production particularly attractive. In addition, live adenovirus vaccines have already been shown to be safe in humans when administered orally in enteric coated capsules. The full-length cDNA of the VP4 of the porcine rotavirus OSU strain was cloned into adenovirus type 5 at a location downstream of the E3 promoter. In this viable recombinant the rotavirus VP4 sequence was substituted for sequences in the adenovirus E3 region. The plaque- purified recombinant (Ad5-OSU VP4), expressed an antigenically authentic VP4 protein as determined by immunoprecipitation. The 84 kDa protein had the same electrophoretic mobility as the native VP4 of OSU rotavirus grown in MA104 cells. Preliminary results from a study in which cotton rats were infected intranasally with live Ad5-OSU VP4 indicated that the animals developed a low level of neutralizing antibodies to the OSU porcine rotavirus.