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。
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.
出处
《环境科学学报》
CAS
CSSCI
CSCD
北大核心
1996年第4期469-474,共6页
Acta Scientiae Circumstantiae
基金
国家自然科学基金
关键词
金属硫蛋白
分离纯化
海洋
重金属污染
red scorpoinfish metallothionein, purification, immunoassay.
