• 关键词：金属硫蛋白  分离纯化,ELISA

• 基金项目：国家自然科学基金(49176276)

• 杨丰
• 国家海洋局第三海洋研究所,厦门 361005

• 陈荣忠
• 国家海洋局第三海洋研究所,厦门 361005

• 徐洵
• 国家海洋局第三海洋研究所,厦门 361005

• 摘要：通过SephadexG-75凝胶过滤和DEAE-SepharoseCL-6B离子交换柱层析,从褐菖的肝脏分离金属硫蛋白,经氨基区组成分析表明:褐菖的金属硫蛋白不含环状氨基酸,半胱氨酸含量达35%,估算分子量为6700道尔顿。等电聚焦显示两条区带,PI分别为4.0和4.2.金属硫蛋白经与牛血清蛋白交联对兔进行免疫,兔抗血清用ProteinA亲和纯化并标记上辣根过氧化物酶,利用酶联免疫吸附测定法(ELISA)对褐菖金属硫蛋白进行分析,测定范围为5-100ng/mL,经诱导和没诱导的肝提取液金属硫蛋白含量分别为450μg/mL和20μg/mL,两者之间差别为20多倍。

• Abstract：Red Scorponifish hepatic metallothionein (MT) was purified by Sephadex G-75 gel filtration and DEAE-Sepharose CL-6B ion-exchange chromatography.The results of amino acid composition analysis showed that the protein contain 35% cysteine, but no aromatic residues.The molecular weight was about 6. 7 KD, and PIs were 4. 0 and 4. 2, respectively. The antibody was raised in rabbits by injection with MT conjugated with BSA, then purified on Protein A beads and labeled with horseradish peroxidase. Quantification of MT using an enzymelinked imino sorbent(ELISA) indicated that the concentrations of MT from induced and noninduced liver extraction were 450 μg/mL and 20μg/mL, respectively.

• Key words：red scorpoinfish metallothionein  purification  immunoassay

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