大肠杆菌过氧化氢酶katE基因的克隆及烟草的遗传转化

Cloning and Construction of Plant Express Vector of Escherichia Coli katE Gene and Transgenic in Tobacco

过氧化氢酶是生物体内主要的抗氧化酶之一,其主要功能是催化细胞内过氧化氢的分解,清除光呼吸、线粒体电子传递以及脂肪酸β-氧化等过程中产生的H2O2,从而使细胞免于遭受过氧化氢的毒害。本实验克隆了大肠杆菌过氧化氢酶katE基因及定位叶绿体的转运肽STP,并构建了植物融合表达载体pB I-STP-katE,重组质粒通过冻融法转入根癌农杆菌GV3101。采用叶盘浸染法转化烟草,获得了部分转基因植株,为进一步研究过氧化氢酶katE基因的功能打下基础。

Catalase is one of the main antioxidant enzymes in Organisms. Catalases can catalyse the decomposition of the cellular hydrogen-peroxide, H2O2 produced in the light breathing, electron transport in mitochondrial and fatty acid β-oxidation process is mainly removed by catalase in the plant, thus protect cells from hydrogen peroxide by the poison. The gene encoding a katE protein and STP transit peptide are cloned from Escherichia coli and Arabidopsis thaliana genome respectively. Plant expression vector of fusion transit peptide of STP and katE gene is constructed, and then introduced into agrobacterium tumefaciens GV3101 via freeze-thaw method. Transgenic tobaccos is ob- tained by leaf-disk transformation. It puts the way for further research katE function.