摘要
旨在研究盐穗木金属硫蛋白Hc MT基因原核表达情况及结合金属离子的能力。从盐穗木中克隆获得的金属硫蛋白(Hc MT)基因亚克隆至p GEX-6p-2原核表达载体上,在大肠杆菌BL21中表达,分离纯化融白蛋白GST-Hc MT,通过原子吸收分光光度法测定融合蛋白GST-Hc MT结合金属离子的量以判断其与金属离子的结合能力。结果显示,诱导表达的融合蛋白GST-Hc MT分子量在34 k D左右,与预期一致,且通过Western blot进一步得到验证;原子吸收结果表明其具有结合Cu^(2+)、Zn^(2+)、Pb^(2+)、Cd^(2+)的能力,融合蛋白结合Cu^(2+)、Zn^(2+)、Pb^(2+)、Cd^(2+)的量分别是对照的25倍、10倍、4倍和2倍,其结合能力大小为Cu^(2+)>Cd^(2+)>Zn^(2+)>Pb^(2+)。
This study aims to evaluate the prokaryotic expression and metal-binding ability of metallothionein protein in Halostachys caspica （ HcMT ） . HcMT cloned from H. caspica was transferred to prokaryotic expression vector pGEX-6p-2 and expressed in Escherichia coli BL21. Then the glutathione S-transferase （ GST ） fusion protein （ GST-HcMT ） was isolated and purified. Using atomic absorption spectrometry, the metal-binding ability of the GST-HeMT was determined by measuring the amount of metal ions it bound with. The results showed that the molecular weight of expressed fusion protein GST-HcMT was about 34 kD, which was as expected and confirmed by Western blot. Furthermore, GST-HcMT bound 25, 10, 4, 2 folds of Cu2＋, Zn2＋, Pb2＋, and Cd2＋separately as GST alone, and the binding ability of GST-HeMT was as follows ： Cu2＋ 〉 Cd2＋ 〉 Zn2＋ 〉 Pb2＋.
出处
《生物技术通报》
CAS
CSCD
北大核心
2016年第4期121-127,共7页
Biotechnology Bulletin
基金
新疆维吾尔自治区高技术研究发展项目(201416102)
关键词
盐穗木金属硫蛋白
原子吸收分光光度法
金属离子结合能力
metallothionein protein from Halostachys caspica （ HcMT ）
atomic absorption spectrometry （ AAS ）
metal-binding ability
