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拟南芥新型花药特异性基因的鉴定、序列分析及表达研究。

Identification, sequence analysis and expression studies of novel anther-specific genes of Arabidopsis thaliana.

作者信息

Rubinelli P, Hu Y, Ma H

机构信息

Cold Spring Harbor Laboratory, NY 11724, USA.

出版信息

Plant Mol Biol. 1998 Jul;37(4):607-19. doi: 10.1023/a:1005964431302.

Abstract

Relatively little is known about pollen development at the molecular level. For the purpose of gaining understanding of the molecular control of pollen development, a number of Arabidopsis cDNA fragments were isolated using subtractive hybridizations. DNA and RNA hybridizations and sequence analyses indicate that we have isolated cDNAs representing 13 genes. Sequences for 8 of these genes are novel, while those for the remaining 5 genes have substantial similarity to genes previously reported as anther- or pollen-specific. RNA in situ hybridizations with 5 genes revealed that four of them are tapetum-specific with differing temporal expression patterns during pollen development and one is pollen-specific within the flower. Sequence analysis of full-length cDNAs showed that one of the novel genes, ATA7, encodes a protein related to lipid transfer proteins. Another gene, ATA20, encodes a protein with novel repeat sequences and a glycine-rich domain that shares a predicted structure with a known cell wall protein. The full-length ATA27 cDNA encodes a protein similar to the BGL4 beta-glucosidase from Brassica napus. The ATA27 protein is predicted to have an ER retention signal and an acidic isoelectric point, suggesting that it may be localized to the ER lumen. This may be a means of compartmentalization from its substrate(s). Our studies demonstrate that subtractive hybridizations can be used to identify previously unknown genes, which should be valuable tools for further study of pollen and anther development and function.

摘要

在分子水平上,人们对花粉发育的了解相对较少。为了深入了解花粉发育的分子调控机制,利用消减杂交技术分离了一些拟南芥cDNA片段。DNA和RNA杂交以及序列分析表明,我们分离出了代表13个基因的cDNA。其中8个基因的序列是新的,而其余5个基因的序列与先前报道的花药或花粉特异性基因有很大的相似性。对5个基因进行RNA原位杂交显示,其中4个基因在花粉发育过程中具有不同的时间表达模式,是绒毡层特异性的,另一个基因在花内是花粉特异性的。全长cDNA的序列分析表明,一个新基因ATA7编码一种与脂质转运蛋白相关的蛋白质。另一个基因ATA20编码一种具有新的重复序列和富含甘氨酸结构域的蛋白质,该结构域与一种已知的细胞壁蛋白具有预测的结构。ATA27全长cDNA编码一种与甘蓝型油菜的BGL4β-葡萄糖苷酶相似的蛋白质。预测ATA27蛋白具有内质网滞留信号和酸性等电点,这表明它可能定位于内质网腔。这可能是将其与底物分隔开的一种方式。我们的研究表明,消减杂交可用于鉴定以前未知的基因,这些基因对于进一步研究花粉和花药的发育及功能应是有价值的工具。

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