重组杆状病毒表达轮状病毒SA11毒株VP4蛋白

VP4 Protein of Simian Rotavirus Strain SA11 Expressed by a Baculovirus Recombinant

从培养的SA11病毒中提取病毒dsRNA作为模板，经逆转录、多聚酶链反应（RT-PCR）获得编码VP4蛋白的全基因，全长2362bp.完整的VP4基因克隆到杆状病毒转移载体pVL-1393中，转染Sf9细胞进行同源重组，用声、杂交法筛选出表达VP4蛋白的重组杆状病毒。表达的VP4蛋白能被抗轮状病毒抗体所识别，表达产量约占总蛋白的10％。用表达的VP4蛋白免疫动物后可产生高水平抗亲本SA11毒株中和抗体。抗体能阻断SA11病毒在MA104细胞上引起的细胞病变（CPE），免疫荧光和Westernblot也证实抗体可特异性识别轮状病毒抗原。结果表明，重组杆状病毒表达的VP4蛋白具有良好的抗原性和免疫原性，可望用VP4基因研制轮状病毒重组疫苗和亚单位疫苗。

The complementary DNA copies of the gene 4 were prepared by reverse transcription and polymerase chain reaction (RT-PCR) from genomic RNA of simian rotavirus strain SA11. A complete VP4 gene was been inserted into a baculovirus intermediate vector pVL-1393 which was under control of the polyhedrin promoter. The outer capsid protein VP4 which was a major neutralization antigen of rotavirus, was expressed in high yield in Spodoptera frugiperda cell line. The VP4 protein expressed was recognized with a hyperimmunized serum directed against the rotavitus by immunoblot assay. Expressed VP4 protein own about 10 percent of the total protiens. The baculovirus recombinant expressed fulllength VP4 protein was used to immunize animals. A high level of neutralizing antibody directed against parentalstrain SA11 virus was developed in the animals. The anti-vp4 antibody could block visible cytopathic effect (CPE) forming of SA11 virus in MA1o4 cells. Immunofluorescence test and Western blot also confirmed that the antibody could recognize specif ic antigen of rotavirus in infectious MA1O4 cel1s. The results suggest that VP4 protein expressed in a recombinant baculovirus has antigenicity and immunogenicity as well. It may be an important component in developing rotavirus recombinant and subunit vaccine.