锦鲤疱疹病毒ORF146基因的克隆、表达及免疫原性的研究

Cloning，Expression and identification of immunogenicity ORF146 Genes of Koi Herpesvirus

关键词:  锦鲤疱疹病毒  ORF146基因  原核表达  免疫原性

Key words:  koi herpes virus  ORF146 gene  prokaryotic expression  immunogenicity

摘要:

为了研究能有效预防锦鲤疱疹病毒(KHV)的疫苗，以KHV-CJ株为模板克隆了锦鲤疱疹病毒ORF146基因，并构建重组质粒PET32a-ORF146，用IPTG诱导表达，SDS－PAGE检测获得含his标签大小为57.4kDa的蛋白条带。蛋白纯化浓缩后免疫小鼠，不同天数采血收集血清用间接ELISA检测抗体水平。结果显示:注射表达蛋白的小鼠血液中可检测到KHV的特异性抗体，在21天左右抗体水平达到最大值。

Abstract:

To study a vaccine which can be used to effectively prevent koi herpes virus,koi herpe svirus ORF146 gene was selected,the recombinant plasmid PET32a-ORF146 was induced by IPTG，SDS-PAGE detection of the protein indicate the size of 57.4kDa bands( contains his tags).Purified and concentrated protein immuned mice,and serum was collected in different days，antibodies level of serum were determined by indirect ELISA．The results show that KHV specificity antibodies can be detected in the blood of the mice injected expressing protein.Antibody level has reached the maximum in about 21 days.