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| 猪肺炎支原体P46基因的克隆及表达 | |
| 引用本文： | 迟灵芝,刘茂军,冯志新,单虎,邵国青.猪肺炎支原体P46基因的克隆及表达[J].江苏农业学报,2008,24(3). |
| 作者姓名： | 迟灵芝 刘茂军 冯志新 单虎 邵国青 |
| 作者单位： | 1. 江苏省农业科学院兽医研究所,江苏,南京,210014;青岛农业大学动物科技学院,山东,青岛,266109 2. 江苏省农业科学院兽医研究所,江苏,南京,210014 3. 青岛农业大学动物科技学院,山东,青岛,266109 |
| 基金项目： | 江苏省自然科学基金，江苏省农业科学院资助项目 |
| 摘 要： | 参考GenBank登录的猪肺炎支原体(Mhp)P46基因序列,利用Primer 5.0软件设计合成引物,通过重叠延伸PCR(SOE-PCR)对猪肺炎支原体P46基因的3个位点进行定点突变,将此突变基因插入到表达载体pET-32a( )中,经IPTG诱导后成功地在大肠杆菌Rosetta(DE3)宿主菌中表达分子量约为63 500的融合蛋白.Western blotting分析表明,该融合蛋白具有较好的免疫原性.为猪肺炎支原体抗体ELISA检测方法的建立奠定了基础. |
| 关 键 词： | 猪肺炎支原体 重叠延伸PCR 定点突变 表达 |
Cloning and Expression of P46 Gene from Mycoplasma hyopneumoniae |
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| CHI Ling-zhi,LIU Mao-jun,FENG Zhi-xin,SHAN Hu,SHAO Guo-qing.Cloning and Expression of P46 Gene from Mycoplasma hyopneumoniae[J].Jiangsu Journal of Agricultural Sciences,2008,24(3). | |
| Authors: | CHI Ling-zhi LIU Mao-jun FENG Zhi-xin SHAN Hu SHAO Guo-qing |
| Abstract: | Based on the gene sequence of Mycoplasma hyopneumoniae(Mhp) P46 reported in GenBank,primers were designed using Primer 5.0.The site-directed mutagenesis was performed by overlap-extension PCR(SOE-PCR) method at three sites of Mycoplasma hyopneumoniae(Mhp) P46.Then the gene was inserted into expression vector pET-32a( ),the fusion protein with 63 500 was induced and expressed in E.coli Rosetta(DE3) with IPTG.Western-blotting analysis proved that the fusion protein have immunogenicity of Mph.It could be useful for the research of the Mph and the method of detecting the Mph antibody. |
| Keywords: | Mycoplasma hyopneumoniae overlap-extension PCR(SOE-PCR) site-directed mutagenesis expression |
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