在昆虫细胞中高效表达蓝舌病毒VP7蛋白作为组特异性诊断抗原

EXPRESSION OF BLUETONGUE VIRUS VP7 IN INSECT CELLS AND ITS APPLICATION FOR DIAGNOSIS OF GROUP SPECIFIC ANTIGEN

对蓝舌病毒（ Ｂ Ｔ Ｖ） 结构蛋白 Ｖ Ｐ７ 作为组特异性诊断抗原进行了研究。将编码 Ｂ Ｔ Ｖ１３主要组特异性抗原 Ｖ Ｐ７ 的 Ｓ７ｃ Ｄ Ｎ Ａ 插入杆状病毒表达载体ｐ Ｆａｓｔ Ｂａｃ１ ，通过同源重组获得了重组杆状病毒ｒｖ Ｂａｃ Ｂ Ｔ Ｖ Ｐ７ 。用此重组病毒感染昆虫细胞获得 Ｖ Ｐ７ 蛋白的高效表达，表达量可占细胞蛋白总量的１２４ ％ 。 Ｖ Ｐ７ 重组抗原与 Ｂ Ｈ Ｋ２１ 细胞培养的抗原具有一致的抗原性，可与不同血清型 Ｂ Ｔ Ｖ 抗体反应，重组抗原的产量是后者的１００ 倍以上。试验证实：所表达的 Ｂ Ｔ Ｖ１３ Ｖ Ｐ７ 抗原能符合组特异性诊断抗原的要求。

In order to develop a novel diagnostic antigen for bluetongue viruses, the cDNA of BTV serotype 13(BTV13) S7, which encodes VP7 and contained main group specific antigen, was inserted into baculovirus expression vector pFastBacl. After homologous recombination, recombinant baculoviruses rvBcBTVP7 were gained. High level expression of VP7 protein was obtained after the infection of rvBacBTVP7 to Sf9 cells. The expressed VP7 could reach up to 15-20% of the total cell proteins. The recombinant VP7 antigen reacted well with BTV sera of different serotypes that was also true to the antigens from BTV infected BHK-21 cells, but the production was 100 times higher than the later, it implies the usefulness of the BTV13 VP7 as a group specific diagnostic antigen.