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| 胸膜肺炎放线杆菌血清10型ApxIA基因的克隆及原核表达 | |
| 引用本文： | 富亮,徐福洲,赵玉军,刘莹.胸膜肺炎放线杆菌血清10型ApxIA基因的克隆及原核表达[J].吉林畜牧兽医,2006,27(4):6-8,11. |
| 作者姓名： | 富亮 徐福洲 赵玉军 刘莹 |
| 作者单位： | 1. 沈阳农业大学畜牧兽医学院,辽宁,沈阳,110161;北京市农林科学院畜牧兽医研究所,北京,100089 2. 北京市农林科学院畜牧兽医研究所,北京,100089 3. 沈阳农业大学畜牧兽医学院,辽宁,沈阳,110161 |
| 基金项目： | 北京市科技新星计划项目;北京市自然科学基金 |
| 摘 要： | Apx毒素在胸膜肺炎放线杆菌致病性和免疫原性方面具有重要作用。为研究ApxI毒素的免疫活性，参照胸膜肺炎放线杆菌血清10型ApxIA基因核酸序列（D16582）设计1对引物．利用PCR自该菌株基因组DNA中扩增出3158bp的ApxIA基因片段，经克隆和序列测定后转入原核表达载体pET-32a中，通过转化BL21（DE3）并在IPTG诱导下进行原核表达，表达出约的融合蛋白，SDS-PAGE和Westernblotting鉴定结果显示表达产物大小约120kDa，且表达产物可与该菌株免疫兔血清发生结合反应。ApxIA基因的克隆和原核表达为研究ApxI毒素的免疫原性以及在胸膜肺炎放线杆菌中的生物学活性奠定了基础。 |
| 关 键 词： | 胸膜肺炎放线杆菌 ApxIA基因 克隆 原核表达 |
| 文章编号： | 1672-2078（2006）04-0006-04 |
| 收稿时间： | 2006-02-28 |
| 修稿时间： | 2006-02-28 |
Cloning and Prokaryotic Expression of the ApxⅠA Gene of Actinobacillus Pleuropneumoniae Serovar 10 |
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| FU Liang,XU Fu-zhou,ZHAO Yu-jun,LIU Ying.Cloning and Prokaryotic Expression of the ApxⅠA Gene of Actinobacillus Pleuropneumoniae Serovar 10[J].Jilin Animal Science and Veterinary Medicine,2006,27(4):6-8,11. | |
| Authors: | FU Liang XU Fu-zhou ZHAO Yu-jun LIU Ying |
| Institution: | 1. College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110161 ; 2. Institute of Animal Science and Veterinary Medicine, Beijing Municipal Academy of Agricultural and Forestry Science, Beijing, 100089 |
| Abstract: | Apx toxin plays a major role in virulence and immunogenecity of A.pleuropneumoniae.In orderto investigate immunogenecity of ApxI toxin,a 3 158 bp fragment of ApxIA gene was amplified from the genomic DNA of the A.pleur-opneumoniae serovar 10 reference strain(D16582).The amplified fragment was cloned into pGEM-T easy vector for se-quence.The expression vector pET-32a was used to express the ApxIA fragment in E.coli BL21(DE3) with the induction of IPTG.An approximately 120 kDa fusion protein appeared in SDS-PAGE electrophoresis,and the expressed product sho-wed immunoreactivity with rabbit serum against A.pleuropneumoniae serovar 10 by Western blotting analysis.Prokaryotic expression of the ApxIA gene offers a potential subunit preparation of ApxI toxin. |
| Keywords: | Actinobailluspleuropneumoniae ApxlA gene Clone Prokaryotic expression |
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