论文部分内容阅读
利用ＲＴ－ＰＣＲ技术克隆了番茄ＡＣＣ氧化酶基因ＬＥＥＴＨＹＢＲ编码区０．９ｋｂ的ｃＤＮＡ片段，经酶切图谱分析和序列分析鉴定后，反向插入到植物表达载体ｐＢｉｎ４３８中，构建了表达ＡＣＣ氧化酶反义ＲＮＡ的二元载体。用农杆菌侵染美洲黑杨叶片，在含卡那霉素的ＭＳ培养基上选择转化子和植株再生，通过ＰＣＲ检测筛选到１６株转基因杨树植株，Ｓｏｕｔｈｅｒｎｂｌｏｔ分析初步确证了外源基因是以单拷贝插入到杨树基因组中；对杨树幼苗乙烯释放量的测定结果表明转基因杨树的乙烯释放量为对照植株的２８％。
The 0.9kb cDNA fragment of tomato ACC oxidase gene LEETHYBR was cloned by RT-PCR. After restriction analysis and sequence analysis, it was inserted into the plant expression vector pBin438 reversely to construct a recombinant expression vector expressing ACC oxidase Binary vector of sense RNA. Agrobacterium tumefaciens infection of Populus tomentosa leaves, MS medium containing kanamycin selection of transformants and plant regeneration, PCR screening of 16 strains of transgenic poplar plants, Southern blot analysis initially confirmed the foreign gene is The single copy was inserted into poplar genome. The results of ethylene production of poplar seedlings showed that the ethylene content of transgenic poplars was 28% of the control plants.