细胞质型APX的纯化、抗体的制备及放线菌素D对APX活性的影响

上海大学学报(自然科学版)

细胞质型APX的纯化、抗体的制备及放线菌素D对APX活性的影响

陈沁1，王伟1，刘友良2

1.上海大学生命科学学院，上海 200444；
2.南京农业大学生命学院，南京 210095

收稿日期:2005-05-25 修回日期:1900-01-01 出版日期:1900-01-01 发布日期:1900-01-01
通讯作者: 陈沁1

Purification of Cytosolic APX from Barley Leaves, Preparation of Antibodies and Effect of AMD on APX Activities

CHEN Qin1,WANG Wei1,LIU You-liang2

1. School of Life Sciences, Shanghai University, Shanghai 200444, China; 
2. Faculty of Life Science, Nanjing Agriculture University, Nanjing 210095, China

Received:2005-05-25 Revised:1900-01-01 Online:1900-01-01 Published:1900-01-01
Contact: CHEN Qin1

摘要/Abstract

摘要：

利用硫酸铵分步沉淀与过柱层析提纯大麦叶片细胞质型APX（Ascorbate peroxidase，抗坏血酸过氧化物酶, EC1.11.1.11），SDSPAGE电泳显示其是分子量约为29 kD的单体.将纯化的APX免疫青紫蓝兔，获取多抗，Western blot证实科品7号、鉴4的细胞质型APX具有免疫交叉性.盐处理后耐盐性不同的品种叶片APX的含量发生变化，APX含量变化的规律与盐胁迫下APX活性变化的规律相似，说明盐胁迫下APX含量的变化是引起其活性变化的主要原因之一.不同浓度的AMD（Actinomycin D，放线菌素D）处理均能抑制盐和非盐胁迫下大麦叶片的APX活性.

关键词: APX, Western blot, 大麦, 放线菌素D(AMD), 抗体, 盐胁迫

Abstract:

Ascorbate peroxidase (APX，EC1.11.1.11) was purified to homogeneity from barley (Hordeum vulgare L.) leaves. APX was determined as a monomer with a molecular mass of about 29 kD by SDS-polyacrylamide gel electrophoresis. Western blot analysis using polyclonal antibodies raised against barley APX revealed that there exists immune cross-reaction of APX between Jian 4 and Kepin 7, and APX contents of two barley cultivars differing in NaCl sensitivity changed in the course of NaCl stress. Changes in APX contents showed the same trends as that of APX activities. APX activities in leaves of barley seedlings were inhibited by Actinomycin D (AMD) at different concentration.

Key words: actinomycin D (AMD)

, antibody, barley, NaCl stress, Western blot,

ascorbate peroxidase (APX)

陈沁;王伟;刘友良. 细胞质型APX的纯化、抗体的制备及放线菌素D对APX活性的影响[J]. 上海大学学报(自然科学版).

CHEN Qin;WANG Wei;LIU You-liang. Purification of Cytosolic APX from Barley Leaves, Preparation of Antibodies and Effect of AMD on APX Activities[J]. Journal of Shanghai University（Natural Science Edition）.