Abstract:
The intact open reading frame (ORF) of foot-and-mouth disease virus (FMDV) Asia I/XJ strain was amplified by RT-PCR and inserted into the transfer vector pVL1393 to generate plasmid pVL-ORF. Bm-N cells were transfected with pVL-ORF and linearized Bm-BacPAK6 DNA, and the recombinant silkworm baculovirus Bm-ORF containing the full ORF of FMDV was obtained. The results of indirect immunofluorescence assay (IFA) showed that Bm-ORF could be expressed efficiently in Bm-N cell. After inoculating the early 5th instar larvae of silkworm, the polyprotein of FMDV could be detected by sandwich ELISA and empty capsid-like particles could be observed under the electron microscope. Expression products from silkworm were used as the antigen to immunize the cattle. The specific antibody was induced in all vaccinated animals. The immunized cattle were challenged with the virulent FMDV Asia I/XJ strain, two of the four cattle were completely protected and clinical symptoms were alleviated and delayed in the others. The results suggest that this strategy might be used to develop the new subunit FMDV vaccine.
摘要:
通过RT-PCR扩增口蹄疫病毒(FMDV)亚洲I/XJ株的完整开放阅读框(ORF),并插入转移载体pVL1393以生成质粒pVL-ORF。用pVL-ORF和线性化的Bm-BacPAK6DNA转染Bm-N细胞,获得了含有FMDV全ORF的重组家蚕杆状病毒Bm-ORF。间接免疫荧光检测(IFA)结果表明,Bm-ORF在Bm-N细胞中可有效表达。接种家蚕5龄早期幼虫后,夹心ELISA可以检测到FMDV的多蛋白,在电子显微镜下可以观察到空的衣壳样颗粒。家蚕的表达产物被用作免疫牛的抗原。在所有接种疫苗的动物中诱导特异性抗体。免疫的牛受到毒力FMDVAsiaI/XJ株的攻击,四头牛中的两头得到了完全保护,其他牛的临床症状得到了缓解和延迟。结果表明,该策略可能用于开发新的FMDV亚单位疫苗。