镉诱导黄瓜金属硫蛋白抗体的制备及纯化

生态学杂志

镉诱导黄瓜金属硫蛋白抗体的制备及纯化

王廷璞;安建平;邹亚丽;马腾;张春成

天水师范学院生化学院，甘肃天水 741001

收稿日期:2008-01-30 修回日期:1900-01-01 出版日期:2008-09-10 发布日期:2008-09-10

Preparation and purification of cadmium-induced antiserum against cucumber-metallothionein.

WANG Ting-pu;AN Jian-ping;ZOU Ya-li;MA Teng;ZHANG Chun-cheng

School of Life Science and Chemistry, Tianshui Normal University, Tianshui 741001, Gansu, China

Received:2008-01-30 Revised:1900-01-01 Online:2008-09-10 Published:2008-09-10

摘要/Abstract

摘要： 用镉诱导黄瓜叶片组织产生金属硫蛋白(MT)，经分离纯化，血清白蛋白偶联后作为抗原。采用背部皮内、皮下和耳缘静脉注射免疫家兔，制备出效价为1∶16的免疫血清。用饱和硫酸铵盐析法和Sephadex G-200柱层析分离纯化IgG。经测定纯化后的IgG蛋白含量为0.9364 mg·ml^-1。用纯化的IgG建立黄瓜Cd-MT的斑点免疫试验检测方法可检测黄瓜Cd-MT的最低含量为200 pg·ml^-1。

关键词: 控释氮肥, 氨挥发, 脲酶活性, pH值, 溶出率

Abstract: The metallothionein (MT) was produced with cucumber leaves under treatment of cadmium. After purification, the MT was coupled with BSA, and then injected subcutaneously or intracutaneously into the back and auricular vein of rabbit to generate antibodies against MT. The measurement showed that the ratio of antibody was 1 to 16. IgG was isolated and purified by (NH₄)₂SO₄saturated precipitation and Sephadex G-200 column. The content of IgG was up to 0.9364 mg·ml^-1 through UV determination. Dot-immunobinding assay revealed that the minimal level of antigen detected by Cd-MT antibody was 200 pg·ml^-1.

Key words: Leaching rate, Controlled-release N fertilizer, Ammonia volatilization, Urease activity, pH value

王廷璞;安建平;邹亚丽;马腾;张春成 . 镉诱导黄瓜金属硫蛋白抗体的制备及纯化[J]. 生态学杂志.

WANG Ting-pu;AN Jian-ping;ZOU Ya-li;MA Teng;ZHANG Chun-cheng . Preparation and purification of cadmium-induced antiserum against cucumber-metallothionein. [J]. cje.

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