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| 胸膜肺炎放线杆菌ApxIA在大肠杆菌中的可溶性表达及纯化 | |
| 引用本文： | 黄良宗,彭启明,王淑敏,张桂红,顾万军.胸膜肺炎放线杆菌ApxIA在大肠杆菌中的可溶性表达及纯化[J].畜牧与兽医,2011,43(11):21-24. |
| 作者姓名： | 黄良宗 彭启明 王淑敏 张桂红 顾万军 |
| 作者单位： | 1. 佛山科学技术学院广东省高校预防兽医学重点实验室,广东佛山528231;华南农业大学兽医学院,广东广州510642 2. 佛山科学技术学院广东省高校预防兽医学重点实验室,广东佛山,528231 3. 华南农业大学兽医学院,广东广州,510642 |
| 摘 要： | 以胸膜肺炎放线杆菌1型参考菌株App-259株基因组DNA为模板,PCR扩增apxIA全基因,构建表达重组质粒apxIA-c2X,并将其转化至大肠杆菌TB1,SDS-PAGE和Western-blot分析表达产物.结果表明:成功构建了apxIA-C2x表达质粒,重组菌在15℃经0.2mmol/L IPTG诱导16 h获... |
| 关 键 词： | 胸膜肺炎放线杆菌 apxIA 融合表达 蛋白纯化 |
Soluble expression and purification of ApxIA of Actinobacillus pleuropneumoniae in Escherichia coli |
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| HUANG Liang-zong,PENG Qi-ming,WANG Shu-min,ZHANG Gui-hong,GU Wang-jun.Soluble expression and purification of ApxIA of Actinobacillus pleuropneumoniae in Escherichia coli[J].Animal Husbandry & Veterinary Medicine,2011,43(11):21-24. | |
| Authors: | HUANG Liang-zong PENG Qi-ming WANG Shu-min ZHANG Gui-hong GU Wang-jun |
| Abstract: | The 3069bp fragment of apxIA was amplified from the reference strain 259 of Actinobacillus pleuropneumoniae by PCR and cloned into the pMAL-c2X vector.The recombinant expression vector was transformed into Escherichia coli TB1 and induced by IPTG for expression.The target protein was purified by Amylose Column.The results showed that the recombinant expression plasmid apxIA-c2X was successfully constructed.The SDS-PAGE and Western-blot analysis showed that the apxIA gene could be expressed efficiently in E.coli TB1 with the induction of 0.2mmol/L IPTG for 16 hours at 15℃.Molecular weight of the fusion protein was about 150ku,and 50% of the target protein was soluble.The purity could reach 95% after purified by Amylose Column.This study applies a basis for the development of diagnostic antigen and subunit vaccine against porcine pleuropneumonia. |
| Keywords: | Actinobacillus pleuropneumoniae apxIA fusion expression protein purification |
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