OsMT-1-4c 基因原核表达载体的构建、表达及重金属抗性研究

    Vector construction, prokaryotic expression and resistance to heavy metals of OsMT-1-4c

    • 摘要: 为研究水稻抗重金属的特性,构建了水稻金属硫蛋白(metallothionein,MT)原核表达载体PGEX4T-3-OsMT-1-4c,并转化到Rossetta 菌株中诱导表达,得到大小为33.643 kDa 的GST-OsMT-1-4c重组蛋白。将得到的沉淀用1×PBS 溶解并在低温下超声破碎,离心并电泳检测,然后纯化得到融合蛋白。并在重金属Cd2+ 胁迫下诱导表达重组载体和空载体菌株,发现能表达GST-OsMT-1-4c 蛋白菌株比表达GST 蛋白菌株抗重金属的能力强。

       

      Abstract: In order to study the resistance of paddy rice to heavy metals, PGEX4T-3-OsMT-1-4c, the prokaryotic expression vector of metallothione for paddy rice was constructed, which was then transformed into strain Rossetta to induce the expression, about 33.643 kDa of GST-OsMT-1-4c fusion protein was obtained. The obtained precipitation was dissolved with 1×PBS and conducted ultrasonic fragmentation at low temperature, followed by the centrifugation and electrophoresis detection, and then was gained through the purification. Furthermore, when the recombinant vector and empty vector strains were induced to express under the stress of heavy metal Cd2+, it was found that the strain to express protein GST-OsMT-1-4c owned stronger resistance to heavy metals than that of protein GST.

       

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