大肠杆菌海藻糖合成酶基因的克隆和表达

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利用Mu转座子细胞内克隆了大肠杆菌海藻糖合成酶otsBA基因,克隆频率为1.45×10-3/Kanr转导子.经遗传互补、酶切和部分序列分析表明otsBA基因位于克隆质粒.亚克隆2.87kb DNA片段至不同拷贝数表达质粒并分别转化大肠杆菌otsBA基因缺失株,转化株恢复在0.5mol /L NaCl培养基上生长的功能,高渗透压诱导实验表明,转化株能够合成克隆基因产物海藻糖,但合成量不受克隆质粒拷贝数影响.海藻糖良好的抗高渗能力可能在农作物育种方面发挥重要作用.为构建含有海藻糖合成酶基因的植物表达载体,并在农杆菌的介导下转入植物,赋予其抗高渗、耐干旱能力奠定了重要的研究基础.
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