| 摘要: |
| 为了研发牛瑟氏泰勒虫病基因工程亚单位疫苗,本实验将p23-IL-18融合基因克隆到pET-28a原核表达载体,构建pET-28a-p23-IL-18重组原核表达质粒,经BamHⅠ和EcoRⅠ双酶切鉴定后在37 ℃用IPTG诱导表达,对表达产物进行SDS-PAGE和Western blotting分析。结果表明,成功构建牛瑟氏泰勒虫pET-28a-p23-IL-18原核表达质粒,目的基因在大肠杆菌中成功获得表达,融合蛋白的分子量约为48ku,并被牛瑟氏泰勒虫阳性血清所识别,具有良好的反应原性。本研究结果为牛瑟氏泰勒虫病基因工程亚单位疫苗的制备奠定了基础。 |
| 关键词: 牛瑟氏泰勒虫 p23-IL-18融合基因 原核表达 |
| DOI： |
| 投稿时间：2014-04-02修订日期：2014-06-15 |
| 基金项目: |
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| Prokaryotic expression of Theileria. Sergenti the p23-IL-18 fusion gene |
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| (Jilin Zhengyue Biological Products Co,Ltd) |
| Abstract: |
| In order to prepare the genetically engineering subunit vaccine of Brovine Theileriosis Sergenti, the p23-IL-18 fusion gene is cloned into prokaryotic pET-28a expression vector .It can construct recombinant prokaryotic expression plasmid pET-28a-p23-IL-18 which is confirmed by restriction endonuclease digestion of BamHI and EcoRI . It can induced expression by IPTG induction at 37 ℃. At last, the protein induced by IPTG was examined by Western blotting. The result showed that the prokaryotic expression plasmid pET-28a-p23-IL-18 was built successfully and the objective gene was successfully expressed in Escherichia coli. The molecular weight of fusion protein weight 48KDa and can be recognized by positive serum against .The result of this research lays the foundations for genetically engineering subunit vaccine of Brovine Theileriosis Sergenti. |
| Key words: Brovine Theileria.Sergenti p23-IL-18 fusion gene Prokaryotic expression |
