| 大肠杆菌中重组GNA蛋白的分离纯化 |
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| 引用本文： | 罗素兰,长孙东亭.大肠杆菌中重组GNA蛋白的分离纯化[J].生物技术,2004,14(2):15-17. |
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| 作者姓名： | 罗素兰 长孙东亭 |
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| 作者单位： | 海南大学海洋学院,海南,海口,570228 |
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| 基金项目： | 国家自然科学基金资助项目 (No.30 2 60 0 66)，海南省自然科学基金资助项目 (30 1 1 1 ) |
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| 摘 要： | 具有特异结合甘露糖基的雪花莲外源凝集素(Galanthus nivalis agglutnin，GNA)具有多种生物活性，在糖蛋白分离、逆转录病毒病和害虫防治等方面有广泛的应用价值。该试验分别采用超声破碎法、冻融裂解法和溶菌酶法破碎重组大肠杆菌细胞后，经尿素或SKL(十二烷基肌氨酸钠水溶液)溶解后，再透析复性获得了在大肠杆菌(E．coli)中高效表达的重组GNA蛋白，并经SDS—PAGE电泳检测GNA的大小、浓度及表达量．通过对诱导表达时间、超声处理的功率、时间、模式、尿素和SKL洗涤浓度，透析条件的优化组合，建立了一套从大肠杆菌细胞中分离重组GNA蛋白的有效方法，为进一步的重组CNA生物活性试验提供了物质基础，
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| 关 键 词： | 大肠杆菌 GNA 蛋白 分离 纯化 重组雪花莲外源凝集素 基因工程 |
| 文章编号： | 1004-311X(2004)02-0015-03 |
| 修稿时间： | 2003年9月17日 |
Isolation and Purification of Recombined GNA Protein in E.coli |
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| LUO Su-lan,ZHANGSUN Dong-ting.Isolation and Purification of Recombined GNA Protein in E.coli[J].Biotechnology,2004,14(2):15-17. |
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| Authors: | LUO Su-lan ZHANGSUN Dong-ting |
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| Abstract: | The mannose-binding lectin from snowdrop (Galanthus nivalis agglutinin:GNA) exhibits several unique properties,which is valuable in isolation of the major glycoprotein,inhibitory activity against retroviruse,development of novel insect contol agents etc.E.coli cells containing mature GNA (Galanthus nivalis agglutinin) gene were broken up by sonication,freeze-thawing and enzyme lysis methods respectively.Recombined GNA protein in E.coli was isolated and purified with urea or SKL dissolving,dialysis renaturation,et al.The mass,concentration and expressive level of recombined GNA were determined by SDS-PAGE.One efficient method of isolating inclusion recombined GNA from E.coli was established through optimizing inducible time,the power,time and mode of sonication,urea and SKL dissolving concentration and dialysis conditions.The results would be the basis for further bioactivity experiment of recombined GNA. |
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| Keywords: | Recombined snowdrop lectin (GNA) Isolation and purification E coli |
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