| 广西眼镜王蛇毒酸性磷脂酶A2—1的表达 |
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| 引用本文： | 舒雨雁 林文珍 庄茂辛 林政炯 吴祥甫 周元聪. 广西眼镜王蛇毒酸性磷脂酶A2—1的表达[J]. 现代临床医学生物工程学杂志, 2003, 9(1): 1-5 |
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| 作者姓名： | 舒雨雁 林文珍 庄茂辛 林政炯 吴祥甫 周元聪 |
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| 作者单位： | [1]广西医科大学蛇毒研究所南宁530021 [2]中国科学院生物物理研究所生物大分子国家重点实验室北京100101 [3]中国科学院上海生命科学院生物化学与细胞生物学研究所上海200031 |
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| 基金项目： | 国家自然科学基金资助项目 (39970 1 74) |
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| 摘 要： | 将广西眼镜王蛇毒酸性磷脂酶A2—1(APLA2—1)基因克隆至表达载体pET28a( )，转化入大肠杆菌BL21，经过诱导，SDS—PAGE检测，重组质粒pET28a—APLA2—1在18 KD有一明显的表达条带，表达产物APLA2—1约占细菌总量30％，并以包涵体的形式存在，将产物变复性后用Sephadex G—75纯化，产物经过SDS—PAGE检测只有单一条带，通过Western—blot检测，证实纯化产物是酸性磷脂酶A2、对表达的APLA2进行酶活性和药理活性的测定、至此我们在大肠杆菌中表达了广西眼镜王蛇毒APLA2，这将为以后对PLA2的结构与功能的研究打下了良好的基础。
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| 关 键 词： | 眼镜王蛇毒 酸性磷脂酶A2－1 表达 复性 活性测定 |
| 文章编号： | 1008-634X(2003)01-0001-05 |
| 修稿时间： | 2002-12-26 |
Expression of Acidic Phospholipase A_2-1 from Guangxi Ophiophagus Hannah (King Cobra) Venom |
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| SHU Yu yan,LIN Wen zhen,ZHUANG Mao xin,et al.. Expression of Acidic Phospholipase A_2-1 from Guangxi Ophiophagus Hannah (King Cobra) Venom[J]. Journal of Modern Clinical Medical Bioengineering, 2003, 9(1): 1-5 |
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| Authors: | SHU Yu yan LIN Wen zhen ZHUANG Mao xin et al. |
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| Abstract: | A cDNA encoding acidic phospholipase A 2-1(APLA 2-1) from Guangxi Ophiophagus Hannah(King Cobra) Venom was inserted into bacterial expression vector pET28a(+) and expressed in E.coli BL21.pET28a-APLA 2-1 was effectively expressed in E.coli BL21.The protein was produced as insoluble inclusion bodies.After partial purification by washing the inclusion bodies with Triton X-100,denaturing and refolding,the renatured recombinant protein was purified by Sephadex G-75.The expressed protein exhibited enzymatic activity and platelet aggregation inhibiting effect.These would provide a good basis for further discussing the relation of structure and function,and the studying of evolution of PLA 2 from snake venom. |
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| Keywords: | Ophiophagus Hannah Venom APLA 2 Expression Activity measure Renaturation |
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