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存在于启动子中的各种顺式作用元件在基因应答逆境的过程中起重要的作用,对这些元件进行适当组合是构建诱导型合成启动子的重要可行途径之一。本研究利用多个逆境应答元件(S盒、D盒、Gst1盒、TCA盒和LURP1基因启动子的-85~-46区)和CaMV35S核心(-46-+8)序列相连,组成一个人工合成启动子PRBS,并构建了GUS为报告基因的转化载体,通过农杆菌介导的遗传转化方法获得其水稻转基因植株。利用T1代转基因水稻株系,通过检测GUS酶活性,研究了PRBS在不同逆境及外源激素作用下的表达特征。结果发现PRBS组成型表达水平低,可不同程度地应答稻瘟病病原菌侵染,机械损伤和外源激素(ABA、SA和MeJA)处理,表明PRBS可作为诱导型启动子用于水稻抗逆基因工程遗传改良。
The various cis-acting elements present in the promoter play an important role in the response of genes to stress, and the proper combination of these elements is one of the most viable ways to construct inducible synthetic promoters. In this study, several stress response elements (S-boxes, D boxes, Gst1 boxes, TCA boxes and LURP1 gene promoter -85 ~ -46) and CaMV35S core (-46- +8) sequences were connected to form a synthetic Promoter PRBS, and constructed the transformation vector of GUS as the reporter gene. The transgenic rice plants were obtained through Agrobacterium-mediated genetic transformation. Using T1 generation transgenic rice lines, the expression characteristics of PRBS under different adverse conditions and exogenous hormones were studied by detecting GUS enzyme activity. The results showed that PRBS was constitutively expressed in low level and responded to pathogen infection of rice blast, mechanical injury and exogenous hormones (ABA, SA and MeJA) to some extent, indicating that PRBS can be used as an inducible promoter for rice anti-stress gene engineering Genetic improvement.