ProDH基因反义表达载体的遗传转化

Genetic Transformation of ProDH Antisense Gene Expression Vector

利用农杆菌介导的叶盘法将ProDH基因反义表达载体分别导入番茄品种‘耐运2000’和烟草中,建立并优化了番茄子叶再生体系。同时采用操作简单、再生周期短、转化效率较高的烟草进行了遗传转化体系的建立。试验获得49个烟草抗性芽和177个番茄抗性芽,用特异性引物对其做PCR扩增验证,共获得9个PCR呈阳性的抗性芽,其中7个烟草抗性芽、2个番茄抗性芽,因此证明外源基因已成功转入番茄的基因组中,为番茄转基因工程的进一步研究奠定基础。

Proline dehydrogenase（ProDH） antisense gene expression vectors were imported to‘Naiyun 2000＇and tobacco by agrobacterium mediated leaf disc method. The tomato leaf regeneration system was established and optimized. Tobacco genetic transformation system was established at the same time which had simpleoperation, short regeneration cycle and high conversion efficiency. The experiment got 49 tobacco resistantbuds and 177 tomato resistant buds. Specific primer PCR amplification was done in the validation, and 9 PCR positive resistant buds were obtained, including 7 tobacco resistant buds and 2 tomato resistant buds. It provedthat the exogenous gene was successfully integrated into the genome of tomato. The experiment could lay the foundation for further tomato transgenic engineer.