Department of Biochemistry, Plant Molecular Genetics Institute, University of Minnesota, 55108, St. Paul, MN, USA.
Plant Mol Biol. 1988 Jul;10(4):311-21. doi: 10.1007/BF00029881.
The primary structure of the α1-tubulin gene of Arabidopsis thaliana was determined and the 5' and 3' ends of its transcript were identified by S1 nuclease mapping experiments. The information obtained was used to (i) predict the amino acid sequence of the α1-tubulin, (ii) deduce the positions of introns within the α1-tubulin gene, and (iii) construct 3' noncoding gene-specific hybridization probes with which to study the pattern of α1-tubulin transcript accumulation in different tissues and at different stages of development. The predicted amino acid sequence of the α1-tubulin has 92% identity with the predicted product of the previously characterized A. thaliana α3-tubulin gene. The coding sequence of the α1-tubulin gene is interrupted by four introns located at positions identical to those of the four introns in the α3 gene. RNA blot hybridization studies carried out with an α1-tubulin gene-specific probe showed that the α1 gene transcript accumulates primarily in flowers, with little transcript present in RNA isolated from roots or leaves. In order to investigate the pattern of α-tubulin gene expression in developing flowers, RNA was isolated from flowers at five different stages of development: flower buds, unopened flowers with pollen, open flowers, flowers with elongating carpels, and green seed pods. RNA blot hybridizations performed with 3' noncoding gene-specific probes showed that the α3 tubulin gene transcript is present in flowers at all stages of development, whereas the α1-tubulin gene transcript could only be detected in RNA from unopened flowers with pollen, open flowers, and flowers with elongating carpels.
拟南芥α1-微管蛋白基因的一级结构已经确定,并通过 S1 核酸酶作图实验鉴定了其转录本的 5' 和 3' 末端。所获得的信息用于:(i) 预测α1-微管蛋白的氨基酸序列,(ii) 推断α1-微管蛋白基因内内含子的位置,以及 (iii) 构建 3' 非编码基因特异性杂交探针,用于研究不同组织和不同发育阶段α1-微管蛋白转录本的积累模式。α1-微管蛋白的预测氨基酸序列与之前表征的拟南芥α3-微管蛋白基因的预测产物有 92%的同一性。α1-微管蛋白基因的编码序列被四个内含子中断,这些内含子的位置与α3 基因中的四个内含子的位置相同。用α1-微管蛋白基因特异性探针进行的 RNA 印迹杂交研究表明,α1 基因转录本主要在花朵中积累,在从根或叶中分离的 RNA 中几乎没有转录本。为了研究发育中的花中α-微管蛋白基因表达的模式,从处于五个不同发育阶段的花中分离 RNA:花蕾、未开放的花粉花、开放的花、有伸长心皮的花和绿色的豆荚。用 3' 非编码基因特异性探针进行的 RNA 印迹杂交表明,α3 微管蛋白基因转录本存在于发育中的花的所有阶段,而α1-微管蛋白基因转录本只能在花粉未开放的花、开放的花和有伸长心皮的花的 RNA 中检测到。