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基因免疫法制备鲤鱼MHC-Ⅱβ多克隆抗体及其部分特性鉴定
发表时间：2010-04-15 浏览量：1814 下载量：623
| 全部作者： | 张彦，杨桂文，安利国 |
| 作者单位： | 山东师范大学生命科学学院 |
| 摘 要： | 提取鲤鱼脾脏总RNA，利用反转录酶-聚合酶链锁反应（reverse transcription-polymerase chain reaction，RT-PCR）方法扩增编码MHC-Ⅱβ链β1，β2结构域的cDNA片段，克隆到载体pcDNA3.1中，构建真核表达载体，鉴定正确后，将重组载体注入小鼠进行基因免疫，4周后取血，制备多克隆抗血清。Western blot结果显示：该抗血清可以与原核表达的MHC-Ⅱβ特异性结合。表明利用基因免疫法成功制备了抗鲤鱼MHC-Ⅱβ的抗血清，为进一步研究鱼类MHC-Ⅱ分子的免疫功能提供了有力工具。 |
| 关 键 词： | 分子生物学；鲤鱼；MHC-Ⅱ；基因免疫；抗体 |
| Title： | Production and characteristic identification of antisera raised against recombinant common carp MHC class Ⅱ β chain by DNA immunity |
| Author： | ZHANG Yan，YANG Guiwen，AN Liguo |
| Organization： | College of Life Science, Shandong Normal University |
| Abstract： | In this paper, total RNA was first extracted from the spleen of common carp, then reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify cDNA fragment encoding β1 and β2 domain of MHC-Ⅱβ chain. This fragment was cloned into pcDNA3.1 and constructed the eukaryotic expression vector. The correct recombinant vector was injected into mice, and after four weeks, the polyclonal antisera were got from these mice. Western blot analysis showed that the antisera could bind with the MHC-Ⅱβ expressed in E.coli specifically. These results indicate that the anti-carp MHC-Ⅱβ serum has been got by gene immunization, providing a powerful tool for further study of fish MHC-Ⅱ molecules. |
| Key words： | molecular biology; Cyprinus carpio L.; MHC-Ⅱ; DNA immunity; antibody |
| 发表期数： | 2010年4月第7期 |
| 引用格式： | 张彦，杨桂文，安利国. 基因免疫法制备鲤鱼MHC-Ⅱβ多克隆抗体及其部分特性鉴定[J]. 中国科技论文在线精品论文，2010，3（7）：670-676. |
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