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利用聚合酶链反应差异显示技术鉴定草莓成熟果实中表达增强的mRNA

Identification of mRNAs with enhanced expression in ripening strawberry fruit using polymerase chain reaction differential display.

作者信息

Wilkinson J Q, Lanahan M B, Conner T W, Klee H J

机构信息

Monsanto Company, Chesterfield, MO 63198, USA.

出版信息

Plant Mol Biol. 1995 Mar;27(6):1097-108. doi: 10.1007/BF00020883.

Abstract

Fruit ripening is a complex developmental process that involves specific changes in gene expression and cellular metabolism. In climateric fruits these events are coordinated by the gaseous hormone ethylene, which is synthesized autocatalytically in the early stages of ripening. Nonclimacteric fruits do not synthesize or respond to ethylene in this manner, yet undergo many of the same physiological and biochemical changes associated with the production of a ripe fruit. To gain insight into the molecular determinants associated with nonclimacteric fruit ripening, we examined mRNA populations in ripening strawberry fruit using polymerase chain reaction (PCR) differential display. Five mRNAs with ripening-enhanced expression were identified using this approach. Three of the mRNAs appear to be fruit-specific, with little or no expression detected in vegetative tissues. Sequence analysis of cDNA clones revealed positive identities for three of the five mRNAs based on homology to known proteins. These results indicate that the differential display technique can be a useful tool to study fruit ripening and other developmental processes in plants at the RNA level.

摘要

果实成熟是一个复杂的发育过程,涉及基因表达和细胞代谢的特定变化。在跃变型果实中,这些事件由气态激素乙烯协调,乙烯在成熟早期自动催化合成。非跃变型果实不以这种方式合成或响应乙烯,但会经历许多与成熟果实产生相关的相同生理和生化变化。为了深入了解与非跃变型果实成熟相关的分子决定因素,我们使用聚合酶链反应(PCR)差异显示技术检测了成熟草莓果实中的mRNA群体。通过这种方法鉴定出了五个表达增强的mRNA。其中三个mRNA似乎是果实特异性的,在营养组织中几乎没有或没有检测到表达。对cDNA克隆的序列分析显示,五个mRNA中的三个与已知蛋白质具有同源性,从而确定了其身份。这些结果表明,差异显示技术可以作为一种有用的工具,在RNA水平上研究植物的果实成熟和其他发育过程。

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