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玉米α-微管蛋白基因有义链和反义链转录本的反向平行表达。

Antiparallel expression of the sense and antisense transcripts of maize alpha-tubulin genes.

作者信息

Dolfini S, Consonni G, Mereghetti M, Tonelli C

机构信息

Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Italia.

出版信息

Mol Gen Genet. 1993 Oct;241(1-2):161-9. doi: 10.1007/BF00280213.

Abstract

In all eukaryotes alpha- and beta-tubulins are encoded by small families of closely related genes and are highly conserved. In Zea mays, at least six different alpha-tubulin coding sequences are known. We describe the isolation from scutellar nodes of the maize inbred line W22 of a clone (CTM5) coding for an alpha-tubulin. On the basis of the 3' end nucleotide sequence, this clone can be assigned to the already reported tua4 gene. Northern analysis demonstrates that CTM5 encodes a 1.5 kb transcript, which is expressed in different tissues of the seed and of the seedling. In order to define the spatial and temporal expression of alpha-tubulin genes, in situ hybridization experiments were performed on these tissues. Unexpectedly, a specific signal was detected with both antisense and sense RNA strands. Temporal and spatial distribution of the two RNAs, however, shows that high levels of the two transcripts are always discordant. In tissues where sense transcripts are highly abundant (embryos at various developmental stages, root tips, pollen grains), the antisense transcripts are expressed in relatively small amounts, while in pericarp, coleoptile, leaves, and scutellar node, where antisense transcripts accumulate, the sense transcript only reaches a very low level. Northern analysis using single-stranded DNA probes confirmed the presence of an antisense transcript of 1.5 kb, prompting speculation about the role of this transcript in the regulation of the expression of alpha-tubulin genes.

摘要

在所有真核生物中,α-微管蛋白和β-微管蛋白由密切相关的基因小家族编码,并且高度保守。在玉米中,已知至少有六个不同的α-微管蛋白编码序列。我们描述了从玉米自交系W22的盾片节中分离出一个编码α-微管蛋白的克隆(CTM5)。根据3'端核苷酸序列,该克隆可归入已报道的tua4基因。Northern分析表明,CTM5编码一个1.5 kb的转录本,它在种子和幼苗的不同组织中表达。为了确定α-微管蛋白基因的时空表达,对这些组织进行了原位杂交实验。出乎意料的是,用反义RNA链和正义RNA链都检测到了特异性信号。然而,两种RNA的时空分布表明,两种转录本的高水平总是不一致的。在正义转录本高度丰富的组织(不同发育阶段的胚、根尖、花粉粒)中,反义转录本的表达量相对较小,而在反义转录本积累的果皮、胚芽鞘、叶片和盾片节中,正义转录本仅达到非常低的水平。使用单链DNA探针的Northern分析证实了1.5 kb反义转录本的存在,这引发了对该转录本在α-微管蛋白基因表达调控中作用的推测。

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