摘要
[Objective] This study aimed to reveal the expression pattern of foreign genes regulated by tomato rbcS3A promoter in transgenic tomato. [Method] Rubisco small subunit promoter rbcS3A was cloned by PCR, fused to the upstream of Gus coding region in a binary vector, and transformed into tomato plants mediated by Agrobacterium. Histochemical staining on PCR positive plants was performed to ana- lyze the expression pattern of the foreign gene regulated by the tomato rbcS3A pro- moter in transgenic tomato. [Result] A total of 15 positive plants were obtained, ac- counting for 33.3%. Histochemical staining showed that the expression level of Gus fusion gene was highest in mature leaf, lower in reproductive organs such as fruit, and not detected in seed. [Conclusion] More positive seedlings were obtained using the modified tissue culture method. Under the control of tomato rbcS3A promoter, exogenous gene highly expressed in transgenic plant leaves, but did not express in seeds and tomato pulp.
[目的]研究番茄rbcS3A启动子调控的外源基因在转基因番茄中的表达模式。[方法]以PCR方法克隆番茄Rubisco小亚基启动子rbcS3A,并构建Rbcs3A-Gus嵌合表达载体p1300RG,将重组载体用农杆菌介导的方法导入到番茄中,最后通过对阳性苗进行Gus活性检测,分析rbcS3A启动子驱动的外源基因在番茄中的表达模式。[结果]试验获得15株PCR阳性苗,阳性苗的获得率为33.3%;Gus活性染色结果显示,番茄rbcS3A控制的外源基因能够在转基因番茄成熟的叶片中高效表达,在果实等生殖器官中表达较弱,在种子中不表达。[结论]调整后的试验方案更有利于获得阳性苗;证实了启动子rbcS3A能够驱动外源基因在番茄营养器官叶片中高效表达而在种子等生殖器官中不表达或表达极低。
