地衣芽孢杆菌耐高温α-淀粉酶基因的克隆、原核表达及其在烟草叶中瞬时表达的研究

来源 :2014年度中国热带作物学会遗传育种专业委员会/热带香料饮料作物专业委员会年会暨学术研讨会 | 被引量 : 0次 | 上传用户:ww447978636
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  为探讨在植物中表达耐高温基因的可行性,本研究从地衣芽孢杆菌中克隆得到耐高温α-淀粉酶基因全长,构建了原核表达载体,转入大肠杆菌中使用IPTG于28℃诱导6小时后,通过SDS-PAGE检测到目的蛋白,分子量约为55KDa,并通过酶活力检测实验证明该蛋白具有耐高温α-淀粉酶活性。同时构建了该基因融合GFP的植物表达载体,通过农杆菌介导瞬时转化烟草下表皮细胞,荧光显微镜观察发现,在烟草下表皮细胞的细胞质、液泡中均有绿色荧光。本研究结果表明所克隆得到的耐高温α-淀粉酶基因可在烟草中成功表达,为后期开展表达耐高温α-淀粉酶的转基因植物的相关研究奠定了实验基础。
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