| 羊种布鲁氏菌M5—90 omp31蛋白原核表达 |
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| 引用本文： | 张红星,陈创夫,王远志,唐利燕.羊种布鲁氏菌M5—90 omp31蛋白原核表达[J].石河子大学学报,2009,27(5):593-596. |
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| 作者姓名： | 张红星 陈创夫 王远志 唐利燕 |
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| 作者单位： | [1]石河子大学生命科学院,石河子832003 [2]石河子太学动物科技学院,石河子832003 [3]石河子大学医学院,石河子832002 |
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| 基金项目： | 973计划前期研究专项，国际科技合作项目，国家自然科学基金，国家自然科学基金项目 |
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| 摘 要： | 克隆、测序布鲁氏菌疫苗株M5—90外膜蛋白基因OMP31，原核表达OMP31并对其检测。从羊种布鲁氏菌M5-90中PCR获得OMP31基因，连接到pBS—T克隆质粒并测序；将测序正确的基因片段克隆人大肠杆菌表达载体pET-28a，进行SDS—PAGE，而后Western—blot检测。结果M5-90疫苗株的OMP3I基因序列与羊种布鲁氏菌参考株16M的同源性为99．03％；外膜蛋白基因OMP31在大肠杆菌表达后能够被Western—blot检测到。结论：表达的布鲁氏菌疫苗株M5-90外膜蛋白OMP31可以被布鲁氏菌特异性血清识别，表现出良好的抗原性，为以后实验室诊断和疫苗的研究做好坚实的上游工作。
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| 关 键 词： | 羊种布鲁氏菌疫苗株M5-90 omp31基因 克隆 测序 Western—blot检测 |
Cloning and Prokaryotic Expression of Omp31 Gene from Brucella M5-90 |
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| Institution: | ZHANG Hongxing , CHEN Chuangfu, WANG Yuanzhi, TANG Liyan ( 1. College of Life Science, Shihezi University, Shihezi 832003, China; 2. College of Animal Scieenee ＆ Technology,Shihezi University, Shihezi 832003, China; 3. College of Medeine, Shihezi University, Shihezi 832003, China) |
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| Abstract: | Brucella melitensis M5-90 vaccine strain OMP31 gene was cloned, sequenced,expressed and detected. OMP31 gene was amplified by PCR,cloned into pBS-T vector and sequenced. The objective gene was recombined into pET-28a vector, expressed and detected by SDS-PAGE and Western-blot. The results showed there was 99.03% identity for OMP31 gene between B. melitensis M5-90 vaccine strain and 16M reference strain and the expressed OMP31 was detected by Western-blot. The conclusion conveyed us that the special serum against bruceUa wild strain could recognize the expressed OMP31 and show it is an immunodominant antigen. This experiment gives us a good upstream work to laboratory diagnosis and study of vaccine. |
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| Keywords: | Brucella M5-90 OMP31 gene cloning sequencing western-blot detection |
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