Laboratory of Plant Resource Sciences, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyamam, Sugitani, Toyama 930–0194, Japan.
Biol Pharm Bull. 2010;33(11):1911-4. doi: 10.1248/bpb.33.1911.
A homology-based cloning strategy yielded two cDNA clones designated Am-cam-1 and Am-cam-2, presumably encoding calmodulin protein from a callus culture derived from the leaf tissues of Aquilaria microcarpa. An appreciable increase in the transcriptional activity of Am-cam-1 was reproducibly observed by exposure of the cell culture to methyl jasmonate, as analyzed by a reverse-transcription polymerase chain reaction. The expression level of the gene also increased when the cells were treated with yeast extract. The transcription of Am-cam-2 was similarly stimulated by the treatment with methyl jasmonate and yeast extract, however, the intensities of the enhanced expression appeared to be lower as compared with that of Am-cam-1. In contrast, Ca(2+)-ionophore A23187 did not show inducing activity for the expression of these two calmodulin genes. These results suggest that Am-cam-1 and Am-cam-2 and their products play important roles in signal transduction processes in methyl jasmonate- and yeast extract-treated cells of A. microcarpa, accompanying the change in the transcriptional activities.
基于同源性的克隆策略产生了两个 cDNA 克隆,分别命名为 Am-cam-1 和 Am-cam-2,它们可能编码来自白木香叶片组织愈伤组织培养物的钙调蛋白蛋白。通过反转录聚合酶链反应分析,发现细胞培养物暴露于茉莉酸甲酯后,Am-cam-1 的转录活性可重复增加。当用酵母提取物处理细胞时,该基因的表达水平也增加。Am-cam-2 的转录也受到茉莉酸甲酯和酵母提取物的类似刺激,然而,与 Am-cam-1 相比,增强表达的强度似乎较低。相比之下,钙离子载体 A23187 对这两个钙调蛋白基因的表达没有诱导活性。这些结果表明,Am-cam-1 和 Am-cam-2 及其产物在白木香细胞中茉莉酸甲酯和酵母提取物处理后的信号转导过程中发挥重要作用,伴随着转录活性的变化。