| 摘要: |
| 为了研究鲤Ⅰ-IFN-γ的免疫效果，根据鲤Ⅰ-IFN-γ的mRNA基因序列（GenBank），构建真核表达质粒pEGFP-N1-Ⅰ-IFN-γ，重组表达质粒与不同剂量的转染试剂EndoFectionTM Max混合后转染鲤鱼肌肉细胞，通过荧光倒置显微镜观察，重组表达质粒pEGFP-N1-Ⅰ-IFN-γ在细胞中成功表达。以多种免疫剂量的重组表达质粒与pIRES-ORF81混合对鲤鱼进行肌肉注射，每周免疫一次，共免疫三次，免疫前采血，收集血清，通过酶联免疫吸附试验（ELISA）检测抗体水平。结果表明：于第三次免疫后两周，抗体水平达到最高，联合免疫与单独免疫核酸疫苗pIRES-ORF81相比，抗体水平明显升高，但差异不显著(P>0.05)。鲤Ⅰ-IFN-γ作为锦鲤疱疹病毒的免疫佐剂具有一定的免疫效果。 |
| 关键词: 鲤I型γ干扰素 锦鲤疱疹病毒 核酸疫苗pIRES-ORF81 免疫效果 |
| DOI： |
| 投稿时间：2016-07-04修订日期：2016-08-05 |
| 基金项目:公益性行业专项-锦鲤疱疹病毒病核酸疫苗的研制及初步应用（106037-2016-2018）；国家现代农业产业体系项目（CARS-46-29) |
|
| Immune Effect of Carp I-IFN-γ as Koi Herpesvirus Nucleic Acid Vaccine Adjuvants |
|
| (JiLin Agricultural University) |
| Abstract: |
| Study on the carp Ⅰ-IFN-γ immune effect, on the basis ofⅠ-IFN mRNA gene sequence Genbank login, The eukaryotic recombinant plasmid pEGFP-N1-Ⅰ-IFN were constructed.Mixing recombinant expression plasmid and transfection reagents in several immune dose transfected carp muscle cells. The expression of recombinant expression plasmid pEGFP-N1-Ⅰ-IFN-γ in carp muscle cells were observed by fluorescent inverted microscope.By using the mixture of recombinant expression plasmidin several immune dose and pIRES - ORF81 injected Carp′s intramuscular and immunized once a week, three times in total. Blood was sampling before injection and serum was collected to detect antibody levels by enzyme-linked immunosorbent assay(ELISA). Two weeks after the third immunization,experiment result showed that antibody titer reach the highest level. Compared with pIRES - ORF81alone immune,antibody level of combined immunization increased obviously, but no significant difference was observed between them(P>0.05). Show that carp Ⅰ - IFN -γ have certain immune effectas koi herpesvirus immune adjuvant. |
| Key words: Ⅰ-IFN-γ koi herpesvirus nucleic acid vaccine pIRES- ORF81 immune effect |
