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玉米(Zea mays L.)中的微管蛋白基因表达。幼苗根发育轴上同工型表达的变化。

Tubulin gene expression in maize (Zea mays L.). Change in isotype expression along the developmental axis of seedling root.

作者信息

Joyce C M, Villemur R, Snustad D P, Silflow C D

机构信息

Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108.

出版信息

J Mol Biol. 1992 Sep 5;227(1):97-107. doi: 10.1016/0022-2836(92)90684-c.

Abstract

Two-dimensional gel/western blot analysis was used to characterize alpha- and beta-tubulin isotype expression along the developmental axis of the maize (Zea mays) seedling primary root. We identified four distinct alpha-tubulin isotypes and a minimum of six beta-tubulin isotypes. This analysis showed differences between the alpha- and beta-tubulin isotypes expressed in rapidly dividing tissue at the root tip and differentiated root tissues proximal to the tip. The alpha 1 and alpha 4 isotypes predominated in samples from immature rapidly dividing tissues such as root tips, whereas in mature tissues such as differentiated root and pollen, alpha 2, alpha 3 and alpha 4 isotypes predominated. The beta 1 and beta 2 isotypes were more abundant in protein samples from root cortex than in samples from the root tip or vascular cylinder. In contrast, the beta 4 and beta 5 isotypes appeared to be more abundant in root tip and vascular cylinder samples than in root cortex samples. Hybridization probes from the 3' non-coding region of six alpha-tubulin cDNA clones were used to quantify the levels of corresponding tubulin transcripts in selected tissues, from embryonic to mature and from largely undifferentiated to highly differentiated. The results from these hybridization experiments showed that all of the alpha-tubulin genes were expressed in all tissues examined, although each gene showed a unique pattern of differential transcript accumulation. A transcript produced from cDNA clone representing the tua5 alpha-tubulin gene was translated in vitro and produced an alpha-tubulin that comigrated with the alpha 2 isotype.

摘要

采用二维凝胶/蛋白质免疫印迹分析来表征沿玉米(Zea mays)幼苗初生根发育轴的α-和β-微管蛋白亚型表达。我们鉴定出四种不同的α-微管蛋白亚型和至少六种β-微管蛋白亚型。该分析显示,根尖快速分裂组织中表达的α-和β-微管蛋白亚型与根尖近端的分化根组织之间存在差异。α1和α4亚型在根尖等未成熟快速分裂组织的样本中占主导地位,而在分化根和花粉等成熟组织中,α2、α3和α4亚型占主导地位。β1和β2亚型在根皮层的蛋白质样本中比在根尖或维管束的样本中更丰富。相反,β4和β5亚型在根尖和维管束样本中似乎比在根皮层样本中更丰富。来自六个α-微管蛋白cDNA克隆3'非编码区的杂交探针用于定量选定组织中相应微管蛋白转录本的水平,这些组织涵盖从胚胎到成熟以及从基本未分化到高度分化的阶段。这些杂交实验的结果表明,所有α-微管蛋白基因在所有检测的组织中均有表达,尽管每个基因都呈现出独特的差异转录本积累模式。从代表tua5α-微管蛋白基因的cDNA克隆产生的转录本在体外进行翻译,并产生了一种与α2亚型共迁移的α-微管蛋白。

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