| 囊尾蚴膜联蛋白32在大肠杆菌中的表达及纯化 |
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| 引用本文： | 郭瀛军, 吴丹, 颜宏利, 孙树汉,.囊尾蚴膜联蛋白32在大肠杆菌中的表达及纯化[J].生物工程学报,2001,17(5):553-556. |
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| 作者姓名： | 郭瀛军 吴丹 颜宏利 孙树汉 |
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| 作者单位： | 第二军医大学医学遗传学教研室, |
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| 基金项目： | 国家高科技研究发展项目基金资助(101-06-05-04). |
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| 摘 要： | 在已获得的囊尾蚴膜联蛋白 (Annexin) 32cDNA的基础上 ,以PCR的方法在cDNA两端加上酶切位点 ,插入原核表达载体pJLA 5 0 3,热诱导表达后 ,外源蛋白大部分以可溶形式表达 ,表达量占菌体总蛋白的 35 %。经(NH4)2SO4分级沉淀、DEAE阴离子交换层析及Sephacry1S 2 0 0凝胶过滤层析得到电泳单一条带 ,Westernblot及抗凝血实验证明表达产物是有生物活性的
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| 关 键 词： | 膜联蛋白 基因表达 分离纯化 大肠杆菌 |
| 文章编号： | 1000-3061(2001)05-0553-04 |
| 修稿时间： | 2001年3月6日 |
High Level Expression and Purification of Cysticercosis cellulose Annexin32 in Escherichia coli |
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| Y J Guo,D Wu,H L Yan,S H Sun.High Level Expression and Purification of Cysticercosis cellulose Annexin32 in Escherichia coli[J].Chinese Journal of Biotechnology,2001,17(5):553-556. |
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| Authors: | Y J Guo D Wu H L Yan S H Sun |
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| Institution: | Department of Medical Genetics, Second Military Medical University, Shanghai 200433, China. guo-yingjun@263.net |
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| Abstract: | In previous work, the cDNA encoding Cysticercus cellulose annexin32 has been cloned. With PCR method, two different restriction Sites were added to each end of the cDNA respectively. Then, the cDNA was inserted into prokaryotic expression vector pJLA-503. After inducing, most foreign protein was expressed in soluble form, which was up to 35% of the total protein of the bacteria. Subsequently, the recombinant Annexin32 was purified with (NH4)2SO4 stepwise precipitation, DEAE-Sepharose FF and Sephacryl S-200 HR chromatography. The final pure protein can been shown as a single band in SDS-PAGE, and the biological activity was verified by Western blot and anticoagulation activity assay. |
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| Keywords: | Annexin gene expression isolation and purification E coli |
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