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  

Isolation and purification of plant protein MAP30 and its DNA-cleaving activity on supercoiled DNA

作　　者：王临旭[1] 孙永涛[1] 杨为松[1] 白雪帆[1] 黄长形[1] 王福祥[1] 

机构地区：[1]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

出　　处：《医学研究生学报》2004年第10期865-867,共3页Journal of Medical Postgraduates

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、Westernblots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 30 0 0 0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 :MAP30对DNA(RNA)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

关 键 词：植物蛋白 分离纯化 超螺旋DNA 

分 类 号：Q946.1[生物学—植物学]