| 布鲁菌外膜蛋白OMP10表达及其抗原性的研究 |
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| 引用本文： | 宫晓炜,周继章.布鲁菌外膜蛋白OMP10表达及其抗原性的研究[J].动物医学进展,2009,30(12). |
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| 作者姓名： | 宫晓炜 周继章 |
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| 作者单位： | 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部草食动物疫病重点开放实验室,农业部兽医公共卫生重点开放实验室,甘肃兰州,730046 |
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| 基金项目： | 家畜疫病病原生物学国家重点实验室基金 |
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| 摘 要： | 设计1对特异性引物对羊布鲁菌16M总DNA进行外膜蛋白omp10的PCR扩增,得到了一个大小为330 bp的目的基因片段(去掉17个氨基酸编码的信号肽),测序证实它与国外报道的羊布鲁菌omp10基因完全一致.将其克隆到表达载体PET-30a中,经酶切、PCR扩增和测序分析,表明重组表达载体构建成功.将此重组质粒转化入大肠埃希菌BL21(DE3)中,IPTG诱导表达,该基因以包涵体的形式在大肠埃希菌中表达,经过包涵体的变性、复性和亲和层析纯化,成功获得大小为14.2 ku的融合蛋白,与理论推测的蛋白分子质量一致;Western blot和间接ELISA试验证明,纯化之后的OMP10重组蛋白可以被布鲁菌阳性血清识别.
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| 关 键 词： | 布鲁菌 OMP10重组蛋白 抗原性 |
Expression and Antigenicity Analysis of the Brucella Outer Membrane Protein 10 |
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| GONG Xiao-wei,ZHOU Ji-zhang.Expression and Antigenicity Analysis of the Brucella Outer Membrane Protein 10[J].Progress In Veterinary Medicine,2009,30(12). |
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| Authors: | GONG Xiao-wei ZHOU Ji-zhang |
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| Institution: | GONG Xiao-wei,ZHOU Ji-zhang (State Key Laboratory of Veterinary Etiological Biology,Key Laboratory of,Key Laboratory of Veterinary Public Health of Ministry of Agriculture,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,730046,China) |
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| Abstract: | An omp10 gene, which was deleted signal peptide, was amplified and a 330 bp brand was obtained by PCR method from total DNA of Brucella melitensis with a pair of specific primers. The product was inserted into an expression vector PET-30a and was confirmed that the expression vector PET-30a/omp10 was correctly constructed after restriction enzyme analysis and PCR. The PET-30a/omp10 was transformed into E.coli BL21(DE3)cell and induced by IPTG. The recombinant protein OMP10 of 14.2 ku in size was expressed i... |
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| Keywords: | Brucella recombinant OMP10 protein antigenicity |
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