论文部分内容阅读
【目的】3-羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3-methylglutaryl coenzyme A reductase,HMGR)和1-去氧-D-木酮糖-5-磷酸还原异构酶(1-deoxy-D-xylulose-5-phosphate reductoisomerase,DXR)是萜类生物合成途径的上游关键酶,构建表达阳春砂HMGR及DXR基因的转基因烟草,以期在模式植物中分析基因的生物学功能。【方法】采用亚克隆的方法构建AvHMGR和AvDXR的重组植物表达载体,电击转化农杆菌EHA105,获得带有目的基因的工程农杆菌;通过农杆菌介导的叶盘转化法转化烟草,筛选转基因植株;采用半定量逆转录—聚合酶链反应(RT-PCR)法分析目的基因在转基因烟草中的表达情况。【结果】构建了重组植物表达载体pBI-AvHMGR和pBI-AvDXR及其工程菌;对转基因烟草的PCR检测结果证实AvHMGR和AvDXR已整合到烟草基因组中;目的基因在部分转基因植株中得到表达。【结论】已构建成功表达阳春砂HMGR及DXR基因的转基因烟草。
【Objective】 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and 1-deoxy-D-xylulose 5- 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) is a key upstream enzyme in the terpenoid biosynthesis pathway. The transgenic tobacco expressing HMGR and DXR genes was constructed to analyze the gene biology in model plants Features. 【Method】 The subclone method was used to construct the recombinant plant expression vector of AvHMGR and AvDXR. The Agrobacterium tumefaciens strain EHA105 was transformed into Agrobacterium tumefaciens by electroporation. Agrobacterium tumefaciens-mediated leaf disc transformation was used to transform tobacco and transgenic plants The expression of the target gene in transgenic tobacco was analyzed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). 【Result】 Recombinant plant expression vectors pBI-AvHMGR and pBI-AvDXR were constructed and their engineered bacteria were constructed. The PCR results of transgenic tobacco confirmed that AvHMGR and AvDXR were integrated into the tobacco genome. The target genes were expressed in some transgenic plants. 【Conclusion】 Transgenic tobacco plants expressing HMGR and DXR genes were successfully constructed.