Ulmasov T, Ohmiya A, Hagen G, Guilfoyle T
Department of Biochemistry, University of Missouri, Columbia 65211, USA.
Plant Physiol. 1995 Jul;108(3):919-27. doi: 10.1104/pp.108.3.919.
Transcriptional activation of the soybean (Glycine max) GH2/4 gene (also referred to as Gmhsp26-A) and increase in abundance of the GH2/4 mRNA (also referred to as pCE54) have been previously shown to occur following treatment of soybean seedlings with auxins, nonauxin analogs, heavy metals, and a variety of other agents. To determine whether the GH2/4 promoter is responsive to an array of different agents, we have analyzed the inducibility of the GH2/4 promoter fused to the beta-glucuronidase reporter gene in transgenic tobacco (Nicotiana tabacum) plants. We have shown that a wide variety of chemical agents induce this promoter in a tissue-specific and concentration-dependent manner. In addition, we have used an affinity-purified antibody raised against recombinant GH2/4 protein to show that the GH2/4 protein increases in response to auxin application and is localized in the cytosol of soybean cells. Recombinant GH2/4 protein can be purified to homogeneity on a glutathione-agarose resin, and the purified protein has glutathione S-transferase activity when assayed with the substrate 1-chloro-2,4-dinitrobenzene.
先前已表明，用生长素、非生长素类似物、重金属及多种其他试剂处理大豆幼苗后，大豆（Glycine max）GH2/4基因（也称为Gmhsp26 - A）的转录激活以及GH2/4 mRNA（也称为pCE54）丰度的增加会发生。为了确定GH2/4启动子是否对一系列不同试剂有反应，我们分析了与β - 葡萄糖醛酸酶报告基因融合的GH2/4启动子在转基因烟草（Nicotiana tabacum）植株中的诱导性。我们已表明，多种化学试剂以组织特异性和浓度依赖性方式诱导该启动子。此外，我们使用针对重组GH2/4蛋白产生的亲和纯化抗体，证明GH2/4蛋白在施加生长素后增加，并定位于大豆细胞的细胞质中。重组GH2/4蛋白可以在谷胱甘肽 - 琼脂糖树脂上纯化至同质，并且用底物1 - 氯 - 2,4 - 二硝基苯测定时，纯化的蛋白具有谷胱甘肽S - 转移酶活性。
