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| 副结核分支杆菌Hed蛋白的抗原性分析及在间接ELISA中的应用 | |
| 引用本文： | 刘倩,唐泰山,廉慧锋,王凯民,张常印,雷治海.副结核分支杆菌Hed蛋白的抗原性分析及在间接ELISA中的应用[J].南京农业大学学报,2011,34(1). |
| 作者姓名： | 刘倩 唐泰山 廉慧锋 王凯民 张常印 雷治海 |
| 作者单位： | 1. 南京农业大学动物医学院,江苏,南京210095 2. 江苏出入境检验检疫局,江苏,南京,210001 3. 山西出入境检验检疫局,山西,太原,030024 |
| 基金项目： | 国家质检总局科技项目(2008IK03) |
| 摘 要： | 从副结核分支杆菌K-10菌株克隆出840 bp的hed基因片段,插入原核表达载体pET-32a(+),转化至Escherichia coliBL21(DE3),在0.8 mmol.L-1 IPTG诱导下进行表达,纯化重组蛋白,将其进行Western-blot分析、小鼠免疫试验,包被ELISA板,建立间接ELISA方法。结果显示:重组蛋白具有良好的抗原性,方阵滴定法确定了间接ELISA方法的抗血清最佳稀释度(100倍)和抗原包被浓度(4.4μg.mL-1),该方法具有较好的特异性和敏感性。 |
| 关 键 词： | 副结核分支杆菌 Hed蛋白 抗原性分析 间接ELISA |
Antigenicity analysis of the Mycobacterium paratuberculosis Hed protein and its application in indirect ELISA |
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| LIU Qian,TANG Tai-shan,LIAN Hui-feng,WANG Kai-min,ZHANG Chang-yin,LEI Zhi-hai.Antigenicity analysis of the Mycobacterium paratuberculosis Hed protein and its application in indirect ELISA[J].Journal of Nanjing Agricultural University,2011,34(1). | |
| Authors: | LIU Qian TANG Tai-shan LIAN Hui-feng WANG Kai-min ZHANG Chang-yin LEI Zhi-hai |
| Institution: | LIU Qian1,TANG Tai-shan2,LIAN Hui-feng3,WANG Kai-min2,ZHANG Chang-yin2,LEI Zhi-hai1(1.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China,2.Jiangsu Entry-Exit Inspection andQuarantine Bureau,Nanjing 210001,3.Shanxi Entry-Exit Inspection and Quarantine Bureau,Taiyuan 030024,China) |
| Abstract: | To detect antigenicity of Hed protein of Mycobacterium paratuberculosis(Map),and establish the ELISA diagnosis,hed gene was cloned from Mycobacterium paratuberculosis K-10,and inserted to pET-32a(+)vector.The recombinant plasmid was transformed into competent Escherichia coli BL21(DE3).The recombinant protein was expressed,purified,and detected by Western-blot and the mice immunologic test.After that it was applied in indirect ELISA.The results showed the recombinant proteins possessed the good antigenicity... |
| Keywords: | Mycobacterium paratuberculosis Hed protein antigenicity analysis indirect ELISA |
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