天花粉蛋白基因导入欧美杨108

Transformation of Trichosanthin Gene to Populus euramericana 108

为获得含有天花粉蛋白(TCS)的转基因欧美杨108植株,进行了植物表达载体的构建以及导入欧美杨108的研究。将北京大学惠赠的含有TCS启动子及结构基因的质粒pT1-3经SacⅠ和BamHⅠ双酶切后与同样双酶切的载体pCambia1301用T4 DNA连接酶相连,转化大肠杆菌,构建植物表达载体pC-tPro-TCS。将构建的植物表达载体转入农杆菌LBA4404,采用农杆菌介导法,将其导入欧美杨108中。结果表明,共得到18个潮霉素抗性愈伤和抗性芽,转化效率为4.3%。抗性体经PCR和PCR-Southern检测,初步确认TCS基因已经导入到欧美杨108中。

In order to obtain transgenic Populus euramericana with Triehosanthin （TCS） gene, a plant expression vector was constructed and TCS gene was transformed into P. euramericana. The plasmid pT1-3 containing TCS promoter and structural gene which was donated by Peking University was digested by Sac I and BamH I , the vector pCambial301 was digested by the same enzymes. The digested and purified TCS genes were cloned into plant expression vector pC-tPro-TCS. The pC- tPro-TCS with TCS gene was successfully transferred into Agrobacterium tumefaciens LBA4404, adapting to Agrobacteriummediated to transform the leaf explants, and 18 resistant shoots were obtained with transformation frequency of 4.3%. The resistant shoots were identified by PCR and PCR-Southern analysis. TCS gene was transferred into P. euramericana 108.