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人肝金属硫蛋白-I_A基因在鱼腥藻中的克隆与表达 总被引:2,自引:0,他引:2
将人工合成的人肝金属硫蛋白(metalothionein,简称MT)-IA基因插入至中间载体pRL-439上强启动子psbA后,再将其与穿梭载体pKT-210相连,得到大肠杆菌-蓝藻穿梭表达载体pKT-MT,用三亲接合转移法将pKT-MT转入丝状体蓝藻-鱼腥藻7120,经链霉素筛选,得到了稳定的转人肝MT-IA基因鱼腥藻.纯化单藻落,液体扩大培养.从鱼腥藻中提取的质粒经Southern印迹分析,确定人肝MT-IA基因已转入鱼腥藻7120中,Western印迹分析表明,金属硫蛋白在转人肝MT-IA基因鱼腥藻中得到了表达.经原子吸收光谱法测定表达量约为700μgMT/g鲜藻,重金属耐受性实验表明,得到了能耐受重金属-镉的转人肝MT-IA基因鱼腥藻,它将在清除水域中重金属污染和医药研究方面发挥重要作用. 相似文献
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Mossbauer spectrum measured for the iron components of photosystem Ⅱ (PS Ⅱ ) particles of spinach is a superposition of 4 doublets. Quadrupole splitting and chemical shifting of doublets I -IV are characteris-tics of proteins with oxidized cytochrome b-559, reduced cytochrome b-559, Fe3 -Q complex and Fe2 -Q complex re-spectively . After the PS II particles are treated with La3 , two doublets of Fe2 disappear and Fe2 is converted into Fe3 , indicating that the reduced cytochrome b-559 has been converted into the oxidized cytochrome b-559, and Fe2 -Q complex into Fe3 -Q complex. The Mossbauer spectrum of PS II particles treated with La3 and Ca2 shows that Ca2 can weaken the inhibitory effect of La3 in part, and a portion of the reduced cytochrome b-559 and Fe-Q complex still exist. 相似文献
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