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利用RNA差异显示技术鉴定拟南芥叶片中响应光胁迫表达的基因。

Identification of genes expressed in response to light stress in leaves of Arabidopsis thaliana using RNA differential display.

作者信息

Dunaeva M, Adamska I

机构信息

Department of Biochemistry and Biophysics, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden.

出版信息

Eur J Biochem. 2001 Nov;268(21):5521-9. doi: 10.1046/j.1432-1033.2001.02471.x.

Abstract

The plant cell responds to light stress by the expression of genes encoding specific stress proteins with possible protective functions. Five genes, the mRNA levels of which increased drastically in response to light stress in mature green leaves of Arabidopsis thaliana were identified and isolated by the differential display technique. These genes were designated Lsr1-Lsr5 (light stress-regulated). Northern blot analysis demonstrated that the transcript level of Lsr1-Lsr5 increased 4- to 17-fold under light stress conditions as compared with leaves incubated at low intensity light. Further analysis of the Lsr1-Lsr5 transcript level under cold stress, heat shock, wounding, desiccation, salt stress, oxidative stress and UV-A irradiation showed that the expression of all five genes was triggered by more than one stress factor. Thus, it was expected that isolated genes encode proteins involved in general stress responses. Homology searches revealed that all of the isolated cDNAs were represented in the GenBank in genomic DNAs and expressed sequence tag (EST) cDNA clones. The Lsr1-Lsr4 genes encoded cytoplasmic proteins with assigned identities, such as ERD15 (early responsive to dehydration), ACT2 (actin 2), LEA14 (late embryogenesis abundant) and MT1a (metallothionein class 1a), respectively. Light stress had not yet been reported to induce or enhance the expression of these genes. The Lsr5 clone encoded a novel protein with high similarity to beta-1,3-galactosyltransferases from human and primates predicted to be located in the Golgi body. Three ORFs homologous to the Lsr5 gene were found on chromosome I and IV of Arabidopsis indicating that a multigene family of these proteins exists in plants. The possible role of Lsr gene products in light stress defences is discussed.

摘要

植物细胞通过表达编码具有潜在保护功能的特定应激蛋白的基因来应对光胁迫。利用差异显示技术,在拟南芥成熟绿叶中鉴定并分离出了五个基因,其mRNA水平在光胁迫下急剧增加。这些基因被命名为Lsr1-Lsr5(光胁迫调节基因)。Northern印迹分析表明,与在低强度光照下培养的叶片相比,Lsr1-Lsr5的转录水平在光胁迫条件下增加了4至17倍。对Lsr1-Lsr5转录水平在冷胁迫、热激、创伤、干燥、盐胁迫、氧化胁迫和UV-A照射下的进一步分析表明,所有这五个基因的表达都由不止一种胁迫因子触发。因此,可以预期分离出的基因编码参与一般应激反应的蛋白质。同源性搜索显示,所有分离出的cDNA在GenBank中的基因组DNA和表达序列标签(EST)cDNA克隆中均有代表。Lsr1-Lsr4基因分别编码具有特定身份的细胞质蛋白,如ERD15(脱水早期响应蛋白)、ACT2(肌动蛋白2)、LEA14(胚胎后期丰富蛋白)和MT1a(1a类金属硫蛋白)。尚未有报道称光胁迫会诱导或增强这些基因的表达。Lsr5克隆编码一种与人和灵长类动物的β-1,3-半乳糖基转移酶高度相似的新蛋白,预计定位于高尔基体。在拟南芥的第一和第四条染色体上发现了三个与Lsr5基因同源的开放阅读框,这表明这些蛋白在植物中存在一个多基因家族。文中讨论了Lsr基因产物在光胁迫防御中的可能作用。

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