【摘要】 利用番茄U3snRNA基因上游启动区和ACC合成酶反义RNA-核酶嵌合基因DNA片段,构建含U3snRNA基因上游启动区-ACC合成酶的反义RNA-核酶嵌合序列的表达载体,重组于植物双元表达载体pGA643中,得到pGU3R.用三亲融合法导入农杆菌LBA4404中,采用叶盘法转化烟草,诱导再生小植株,获得了卡那霉素的抗性植株.提取抗性植株总DNA,通过PCR、PCRSouthern杂交检测并分别用启动区序列和ACC合成酶的反义RNA-核酶嵌合序列作探针,通过Southern杂交检测,已筛选出整合有外源基因的转化植株.为进一步研究U3snRNA上游启动区增强反义RNA-核酶基因的表达奠定了基础.更多还原

【Abstract】 The plant expression vector pGU3R, carrying a tomato U3snRNA promoter and ACC synthase antisense RNA-Ribozyme chimeric DNA sequences, has been constructed.pGU3R was introduced into Agrobacterium tumefaciens by triparental mating. Tobacco leaf discs were transformed via Agrobacterium tumefaciens mediated procedure.The transgenic tobacco plantlets formed in induction medium and plants resistant to kanamycin were obtained.PCR and Southern blot analyses confirmed that U3snRNA promoter and ACC synthase antisense RNA-Ribozyme chimeric DNA sequence have been integrated into the genome of tobacco.更多还原