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编码种子特异性生物素化蛋白SBP65的豌豆基因的克隆与表达。

Cloning and expression of the pea gene encoding SBP65, a seed-specific biotinylated protein.

作者信息

Dehaye L, Duval M, Viguier D, Yaxley J, Job D

机构信息

Laboratoire Mixte CNRS/Rhône-Poulenc (Unité associée au Centre National de la Recherche Scientifique, U.M. 41), Rhône-Poulenc Agrochimie, Lyon, France.

出版信息

Plant Mol Biol. 1997 Nov;35(5):605-21. doi: 10.1023/a:1005836405211.

DOI:10.1023/a:1005836405211
PMID:9349282
Abstract

Besides biotin-dependent carboxylases, which play key roles in basic metabolism, SBP65 (seed biotinylated protein of 65 kDa of apparent molecular mass), an atypical biotinylated protein, has been described in pea plants. This seed-specific protein is devoid of any carboxylase activity, and shares many physiological and molecular features with late embryogenesis-abundant (Lea) proteins. In a first step toward understanding the role of this peculiar protein, we have demonstrated the role of abscisic acid (ABA) and of the osmotic environment on its expression using northern blot analysis from immature embryos cultured in vitro and germinating mature seeds. Moreover, the cloning and characterization of its gene (referred to as sbp gene) allowed us to define various potential cis-acting elements within the promoter region to account for the observed strict seed-specific expression. The results described in this paper are consistent with a model in which ABA regulates, at least in part, expression of this gene. However, unlike most lea genes, ABA regulation of the sbp gene seems to occur in a very restricted fashion, being confined only to particular stages of embryo development. Such a strict spatial and temporal expression pattern is dependent on the osmotic environment of the developing embryos and on tissue-specific factors, presumably preventing biotin depletion in cells requiring this essential cofactor for basic metabolic activity.

摘要

除了在基础代谢中起关键作用的生物素依赖性羧化酶外,豌豆植株中还发现了一种非典型的生物素化蛋白——SBP65(表观分子量为65 kDa的种子生物素化蛋白)。这种种子特异性蛋白没有任何羧化酶活性,并且与胚胎后期丰富(Lea)蛋白具有许多生理和分子特征。为了初步了解这种特殊蛋白的作用,我们通过对体外培养的未成熟胚和萌发的成熟种子进行Northern印迹分析,证明了脱落酸(ABA)和渗透环境对其表达的影响。此外,对其基因(称为sbp基因)的克隆和表征使我们能够确定启动子区域内的各种潜在顺式作用元件,以解释所观察到的严格的种子特异性表达。本文所述结果与ABA至少部分调节该基因表达的模型一致。然而,与大多数lea基因不同,sbp基因的ABA调节似乎以一种非常受限的方式发生,仅局限于胚胎发育的特定阶段。这种严格的时空表达模式取决于发育中胚的渗透环境和组织特异性因子,大概是为了防止在需要这种基本代谢活性必需辅助因子的细胞中生物素耗竭。

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