烟草Ⅰ类几丁质酶和β-1,3葡聚糖酶基因茎尖转化太子参的研究

Introduction of Tobacco I-Chitinase and I-β-1,3-Glucanase Gene into pseudostellaria heterophylla following Transformation with Shoot Apex

以烟草为材料,对经过修饰的烟草Ⅰ类几丁质酶和Ⅰ类-β1,3-葡聚糖酶cDNA进行了克隆和序列分析,然后构建到一个双-T植物表达载体130上。用太子参无菌苗的茎尖分生组织为受体,采用农杆菌介导法进行转化。感染小苗经共培养后用除草剂筛选,然后对除草剂抗性植株进行分子检测,获得了转基因植株。

The modified I-chitinase and Ⅰ-1, 3-glucanase from the Nicotiana tabacumcv NC89 were cloned and sequenced. Then they were cloned into a plant double-T expression vector. The apical meristems of Pseudostellaria heterophylla plantlets coming from sterilized seeds were used as the receptor of Agrobacterium-mediated transformation. After cocultivation, the plantlets were transferred to selected media. Fertile transgenic plants were obtained from herbicide-resistant ones and confirmed by PCR and PCR-southern assay.