【摘要】 用冻融法将含有绿色荧光蛋白基因的质粒ｐＧＬ２－ＧＳ导入农杆菌菌株ＬＢＡ４４０４ （ｐＴＯＫ２３３），两个质粒发生同源重组，形成一个新的二元载体ｐＴＯＫ２３３－ＧＳ．农杆菌菌株ＬＢＡ４４０４ （ｐＴＯＫ２３３－ＧＳ） 经叶盘法转化烟草，筛选具有卡那霉素抗性的愈伤组织，诱导成苗．ＰＣＲ 扩增发现，绿色荧光蛋白基因在９０％ 的再生植株中存在．在荧光显微镜下，使用蓝色激发光观察再生植株的徒手切片和压片发现，８０ ％ 以上的再生植株都能不同程度地发出绿色荧光．多数植株的根尖和叶脉都发光，一些植株的气孔、叶表皮或叶绒毛也发光更多还原

【Abstract】 Plasmid pGL2 GS containing gfp was constructed and introduced into A. tumenfaciens strain LBA4404(pTOK233). A new strain LBA4404(pTOK233 GS) was generated by homologous recombination between plasmids pTOK233 and pGL2 GS. Discs of tobacco leaves were transformed with LBA4404(pTOK233 GS) strain. Calli with kanamycin resistance and GUS activity were selected and induced to regenerate tobacco plants. PCR amplification showed that 90% of the Kan R plants contain gfp. The observation of gfp expression in tobacco plants with slides was carried out under fluorescent microscope upon excitation of blue light. It was found that more than 80% of plants shone green fluorescence of various strengthes. Most of them shone green fluorescence at bundle and root tips. In a few plants, green fluorescence could also be seen at air pore, epiderm or tomentum.更多还原