锦鲤疱疹病毒-CJ株ORF126基因的原核表达及免疫试验研究

Immunity test research and prokaryotic expression of ORF126 gene of koi herpesvirus-CJ strain

为了研究能有效预防锦鲤疱疹病毒的疫苗,选取了锦鲤疱疹病毒膜糖蛋白基因ORF126并构建重组质粒p ET32aORF126,SDS-PAGE检测获得含his标签大小为49.9 k Da的蛋白条带。蛋白纯化后免疫鲤鱼,采集不同天数血清,用间接ELISA检测抗体水平。结果显示,用10μg/尾、20μg/尾和40μg/尾的蛋白免疫后均可检测到KHV的抗体,3者之间的抗体水平差异不显著(P>0.05)。

To study a vaccine which can effectively prevent Koi herpes virus, Koi herpes virus ORF136 gene was selected. The recombinant plasmid PET32a-ORF126 was construeted, and SDS-PAGE detection of the protein indicated that the size was 49.9 kDa （Contains his Tags）. Purified protein was used to immunize earps, and serum samples were collected in different days Antibody levels of serum samples were determined by indirect ELISA. The results showed that health koi could induce the production of KHV antihodies after proteins at 10 μg/fish, 20 μg/fish and 40 μg/fish were iutramuseularly injected into heahh kois, respectively. After immunized with 10 μg, 20 μg, and 40 μg proteins, the KHV antibodies could be detected, and the differences were not significant （P〉0.05）.