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首页> 外文期刊>Journal of Biotechnology >Stress regulated expression of the GUS-marker gene (uidA) under the control of plant calmodulin and viral 35S promoters in a model fruit tree rootstock: Prunus incisaxserrula
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Stress regulated expression of the GUS-marker gene (uidA) under the control of plant calmodulin and viral 35S promoters in a model fruit tree rootstock: Prunus incisaxserrula

机译：在植物钙调蛋白和病毒35S启动子的控制下，在模型果树砧木Prunus incisaxserrula中，GUS标记基因（uidA）的胁迫调控表达

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The fact that calmodulin genes (CaM) are tightly associated with the Cau2u+ regulatory pathway, as well as their putative role in plant defence against pathogens, indicate a potential use of alternative plant promoters to express genes of interest in specific tissues or developmental stages. To study the expression level of the apple CaM promoter, 981bp sequences upstream were fused to the uidA gene, introduced into cherry and compared with a 35S-GUS construct. Transgene copy number and transgenic expression levels were analysed using Southern blot, Western blot and RT-PCR techniques. Transcription levels were assessed by GUS fluorometry, histochemistry and real-time PCR techniques in leaves of plantlets grown in vitro under various abiotic stresses like low- and high-temperature, salicylic acid and wounding, harvested after 0, 0.5, 1, 2, 4, 10, 24 and 72h. Histochemical analyses showed staining only in veins and petioles of CaM-GUS lines, while in 35S-GUS plants staining extended to the entire leaf. Furthermore, real-time qPCR data indicate that both promoters are differently regulated by various stresses. Obtained results suggest that the selected apple CaM promoter responsible for the expression of a gene in vascular tissues may offer interesting perspectives for plant defense programs.
机译：钙调蛋白基因（CaM）与Cau2u +调节途径紧密相关的事实，以及它们在植物防御病原体中的推定作用，这一事实表明，可能使用替代植物启动子在特定组织或发育阶段表达目的基因。为了研究苹果CaM启动子的表达水平，将上游981bp序列与uidA基因融合，引入到Cherry中，并与35S-GUS构建体进行比较。使用Southern印迹，Western印迹和RT-PCR技术分析转基因拷贝数和转基因表达水平。通过GUS荧光法，组织化学和实时PCR技术评估了在0、0.5、1、2、4、0、0.5、0.5的低温和高温下收获的各种非生物胁迫（如低温和高温）下体外生长的幼苗叶片中的转录水平，10、24和72h。组织化学分析显示仅在CaM-GUS系的静脉和叶柄中有染色，而在35S-GUS植物中，染色扩展到整个叶片。此外，实时qPCR数据表明两个启动子受各种压力的调控不同。所得结果表明，所选择的苹果CaM启动子负责在血管组织中表达基因，可能为植物防御计划提供有趣的视角。

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