Sun M, Giambiagi S, Burrone O
Institute of Medical Biology, CAMS, Kunming.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1997 Feb;19(1):48-53.
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 proteins. The baculovirus recombinant expressed full-length 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 MA104 cells. Immunofluorescence test and Western blot also confirmed that the antibody could recognize specific antigen of rotavirus in infectious MA104 cells. 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.
通过逆转录和聚合酶链反应（RT-PCR）从猿猴轮状病毒SA11株的基因组RNA制备基因4的互补DNA拷贝。将完整的VP4基因插入受多角体蛋白启动子控制的杆状病毒中间载体pVL-1393中。轮状病毒的主要中和抗原外衣壳蛋白VP4在草地贪夜蛾细胞系中高产表达。通过免疫印迹分析，表达的VP4蛋白可被针对轮状病毒的超免疫血清识别。表达的VP4蛋白约占总蛋白的10%。杆状病毒重组体表达的全长VP4蛋白用于免疫动物。动物体内产生了针对亲本SA11病毒的高水平中和抗体。抗VP4抗体可阻断MA104细胞中SA11病毒可见细胞病变效应（CPE）的形成。免疫荧光试验和蛋白质印迹法也证实该抗体可识别感染性MA104细胞中轮状病毒的特异性抗原。结果表明，重组杆状病毒表达的VP4蛋白具有抗原性和免疫原性。它可能是开发轮状病毒重组疫苗和亚单位疫苗的重要组成部分。
