germin基因的克隆及其在烟草中的表达

摘要：克隆了具有草酸氧化酶活性的小麦(Triticum aestivum )Germin蛋白的基因，构建了植物表达载体并用根癌农杆菌(Agrobacterium tumefaciens )介导法转化烟草。Southern杂交和酶活性检测表明，germin基因在转基因烟草(Nicotiana tabacum )中得到整合、表达，并正确组合成具有草酸氧化酶活性的同源六聚体蛋白。离体实验表明，外源germin基因的表达提高了转基因烟草离体叶片对草酸的耐受性。

Cloning of germin Gene and Its Expression in Tobacco

Abstract: Oxalic acid was produced by several plant pathogenic fungi and thought to ploy a primary role in the pathogenicity of fungal species, including the wide host-range pathogen Sclerotinia sclerotiorum. In the current work, a germin gene from wheat (Triticum aestivum )was cloned. A plant expression vector containing the germin gene was constructed by CaMV 35S promoter, which was used for Agrobacterium tumefaciens-mediated transformation of tobacco(Nicotiana tabacum ). Southern blotting assay indicated that the target gene was integrated into tobacco genome. Enzymatic activity assay of oxalate oxidase in transgenic tobacco showed that the germin gene was expressed, which resulted in the formation of homohexamer with enzymatic activity correctly, and the transgenic plants were tolerant of the exogenously supplied oxalic acid.