| 苦瓜子抗人免疫缺陷病毒蛋白30的分离纯化及其切割超螺旋DNA的活性研究 |
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| 引用本文： | 王临旭,孙永涛,杨为松,白雪帆,黄长形,王福祥. 苦瓜子抗人免疫缺陷病毒蛋白30的分离纯化及其切割超螺旋DNA的活性研究[J]. 医学研究生学报, 2004, 17(10): 865-867 |
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| 作者姓名： | 王临旭 孙永涛 杨为松 白雪帆 黄长形 王福祥 |
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| 作者单位： | 第四军医大学唐都医院解放军感染病诊疗中心,陕西西安,710038 |
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| 摘 要： | 目的：分离纯化苦瓜子抗人免疫缺陷病毒(HIV)蛋白30(MAP30)并对其切割超螺旋DNA的活性作初步分析。方法：采用离子交换层析及凝胶过滤层析等方法从苦瓜子中分离纯化MAP30，并经SDS-PAGE、Western blots等鉴定，将质粒pUC18与不同浓度MAP30孵育，进行琼脂糖电泳分析其切割DNA的活性。结果：得到相对分子质量为30000的目的蛋白MAP30，证实其具有使超螺旋DNA断裂为缺口环状及线状DNA的活性。结论：MAV30对DNA(RNA)的切割作用可能是其发挥生物学效应的主要机制之一。
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| 关 键 词： | 植物蛋白 分离纯化 超螺旋DNA |
| 文章编号： | 1008-8199(2004)10-0865-03 |
| 修稿时间： | 2004-03-17 |
Isolation and purification of plant protein MAP30 and its DNA-cleaving activity on supercoiled DNA |
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| WANG Lin-xu,SUN Yong-tao,YANG Wei-song,BAI Xue-fan,HUANG Chang-xing,WANG Fu-xiang. Isolation and purification of plant protein MAP30 and its DNA-cleaving activity on supercoiled DNA[J]. Bulletin of Medical Postgraduate, 2004, 17(10): 865-867 |
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| Authors: | WANG Lin-xu SUN Yong-tao YANG Wei-song BAI Xue-fan HUANG Chang-xing WANG Fu-xiang |
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| Abstract: | Objective: To isolate and purify plant protein MAP30 (momordica anti-HIV protein of 30000) from the seeds of Momordica charantia and observe its DNA-cleaving activity on supercoiled DNA. Methods:MAP30 was purified and identified with ion exchange chromatography, gel permeation chromatography, SDS-PAGE and Western blots. The DNA-cleaving activity was analyzed by electrophoresis in 1% agarose after Supercoiled plasmid pUC18 as substrate treated with MAP30 of various concentrations. Results: A single band with a molecular mass corresponding to 30 000 was obtained for MAP30. The conversion of Supercoiled plasmid pUC18 to relaxed (nicked) and linear forms was demonstrated. Conclusion:MAP30 is a 30 000 plant protein. The DNA-cleaving activity may play a key role in the mechanism of its biological effects |
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| Keywords: | Plant protein Isolation and purification Supercoiled circuler DNA |
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