甘蔗C_4磷酸烯醇式丙酮酸羧化酶基因转化烟草研究初报

Preliminary Study of Genetic Transformation of Tobacco with Sugarcane C_4 Phosphoenolpyruvate Carboxylase Gene

将全长6.8 kb含所有内含子及部分5’侧端和3’末端的甘蔗C4磷酸烯醇式丙酮酸羧化酶(PEPC)基因插入植物目的基因载体pBI121,经农杆菌介导转化获得转基因烟草,PCR检测和dot southern分析证实外源基因已整合进入烟草基因组中。甘蔗C4Ppc基因导入烟草后其叶片PEPC活性有所提高,但提高较小。因此,有必要继续进行克隆目的基因的5’侧端和3’末端获得其完整序列后用于C3作物的遗传转化研究。

The C4 phosphoenolpyruvate carboxylase gene of 6.8 kb long from sugarcane which includes all introns, partial 5'flanking and partial 3'ending sequence was inserted into pBI121. Transgenic tobacco plants mediated by Agrobacterium tumefaciens were obtained. PCR test and dot Southern analysis showed that the sugarcane C4 Ppc had been integrated into tobacco genome. The PEPC activities in the leaves of these T1 transgenic plants were much higher than those of the non -transformed plants, but lower than other C4 Ppc transgenic plants. Therefore, it is necessary to get intact sugarcane C4 Ppc by cloning the 5'flanking and 3'ending sequence for transformation of C3 crops.