捻转血矛线虫免疫调节型DNA疫苗的构建及在山羊体内表达研究

Construction of Immune Regulatory DNA Vaccines for Haemonchus contortus and Their Expressions in Goats

为构建免疫调节型DNA疫苗,将山羊IL-2基因分别与捻转血矛线虫H11-1、H11-2和H11-3基因串联构建融合表达载体pcDNA/IL-2-H11-1、pcDNA/IL-2-H11-2和pcDNA/IL-2-H11-3。为检测疫苗在山羊体内的表达情况,将纯化的疫苗质粒肌肉注射山羊后取注射部位肌肉组织,于首次免疫7 d后和二次免疫10 d后分别用RT-PCR和Western Blot等方法检测H11抗原的转录和翻译情况。结果发现H11基因和IL-2基因均能在注射部位肌肉获得转录和翻译。

To construct immune regulatory DNA vaccines for Haemonchus contortus, three gene fragments encoding parts of antigen H11, named as H11-1, H11-2 and H11-3, were ligated with goat interleukin-2 gene one by one, and the fusions were expressed in vector pcDNA. In order to analyze the expression of the immune regulatory DNA vaccines in vivo, lambs were administered with the mixture of pcDNA/IL-2-H11-1, pcDNA/IL-2-H11-2 and pcDNA/IL-2-H11-3. Goat muscles were taken from the injected site, the transcriptions and translations of vaccines were tested by RT-PCR and Western Blot analysis, seven days post primary immunization and ten days after boosting. The results showed that immune regulatory DNA vaccines carrying Hll and IL-2 genes were expressed in goat muscle successfully.