Russell D A, Sachs M M
Department of Biology, Washington University, St. Louis, Missouri 63130.
Plant Cell. 1989 Aug;1(8):793-803. doi: 10.1105/tpc.1.8.793.
Two cDNA clones for maize cytosolic glyceraldehyde-3-phosphate dehydrogenase are described. One is about 97% similar in coding capacity to a previously published clone [Brinkmann et al. (1987). J. Mol. Evol. 26, 320-328], while the other shows only 88% similarity. Evidence points toward the three cDNAs being the products of three genes, to be called Gpc1, Gpc2, and Gpc3. When the least similar clone, corresponding to Gpc3, was used to analyze RNA gel blots, anaerobic treatment for 6 hours induced RNA accumulation in the shoots 15.6-fold, while a 1-hour shift from 28 degrees C to 40 degrees C increased accumulation 5.1-fold. Roots had a higher basal level of expression, leading to a 6.0-fold anaerobic induction, and a 2.4-fold heat stress induction. RNA gel blot analysis using the clone corresponding to Gpc2 showed decreased RNA accumulation within 6 hours of anaerobiosis, while analysis with the previously published clone, corresponding to Gpc1, showed a decrease within 24 hours. Neither Gpc1 nor Gpc2 showed heat stress induction, while some other known anaerobic genes did. Through the use of hybrid selection, in vitro translation, and immune precipitation, the relative expression of the three genes is shown. The role of the observed changes in gene expression is discussed in relation to stress physiology.
本文描述了两个玉米胞质甘油醛-3-磷酸脱氢酶的cDNA克隆。其中一个克隆的编码能力与之前发表的克隆[Brinkmann等人(1987年)。《分子进化杂志》26卷,320 - 328页]约97%相似,而另一个仅显示88%的相似性。有证据表明这三个cDNA是三个基因的产物,分别命名为Gpc1、Gpc2和Gpc3。当使用与Gpc3对应的相似性最低的克隆来分析RNA凝胶印迹时,6小时的厌氧处理使茎中RNA积累增加了15.6倍,而从28℃到40℃的1小时温度转变使积累增加了5.1倍。根的基础表达水平较高,导致厌氧诱导增加6.0倍,热应激诱导增加2.4倍。使用与Gpc2对应的克隆进行RNA凝胶印迹分析显示,厌氧处理6小时内RNA积累减少,而使用与Gpc1对应的先前发表克隆进行分析显示,24小时内RNA积累减少。Gpc1和Gpc2均未显示热应激诱导,而其他一些已知的厌氧基因则显示有热应激诱导。通过杂交选择、体外翻译和免疫沉淀,展示了这三个基因的相对表达情况。文中还讨论了观察到的基因表达变化在应激生理学方面的作用。