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植物RNA伴侣在大肠杆菌中的异位表达赋予多种胁迫耐受性

Ectopic Expression of Plant RNA Chaperone Offering Multiple Stress Tolerance in E. coli.

作者信息

Jabeen Bushra, Naqvi S M Saqlan, Mahmood Tariq, Sultana Tasawar, Arif Madiha, Khan Fariha

机构信息

Department of Plant Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan.

Department of Biochemistry, PMAS Arid Agriculture University, Rawalpindi, 46300, Pakistan.

出版信息

Mol Biotechnol. 2017 Mar;59(2-3):66-72. doi: 10.1007/s12033-017-9992-z.

Abstract

Members of the plant glycine-rich RNA-binding proteins (GR-RBPs) family have been reported in flowering, development, circadian rhythms, biotic and abiotic stresses. Particularly, GR-RBPs are reported to function as RNA chaperones, promoting growth and acclimation during cold shock. It is indispensable to further question the efficacy and mechanism of GR-RBPs under various environmental strains. Monitoring the expression of stress-regulated proteins under stress conditions has been a beneficial strategy to study their functional roles. In an effort to elucidate the NtGR-RBP1 function, stress markers such as salinity, drought, low temperature and heat stresses were studied. The NtGR-RBP1 gene was expressed in E. coli followed by the exposure to stress conditions. Recombinant E. coli expressing NtGR-RBP1 were more tolerant to stresses, e.g., salinity, drought, cold and heat shock. Recombinants exhibited higher growth rates compared to control in spot assays. The tolerance was further confirmed by monitoring the growth in liquid culture assays. Cells expressing NtGR-RBP1 under salt (500 mM NaCl), drought (20% PEG), cold (4 and 20 °C) and heat stresses (50 °C) had enhanced growing ability and better endurance. Our study supports the notion that the protective role of NtGR-RBP1 may contribute to growth and survival during diverse environmental stresses.

摘要

植物富含甘氨酸的RNA结合蛋白(GR-RBPs)家族成员已在开花、发育、昼夜节律、生物和非生物胁迫方面有相关报道。特别地,据报道GR-RBPs可作为RNA伴侣,在冷休克期间促进生长和适应。进一步探究GR-RBPs在各种环境胁迫下的功效和机制是必不可少的。在胁迫条件下监测胁迫调节蛋白的表达一直是研究其功能作用的有益策略。为了阐明NtGR-RBP1的功能,研究了盐度、干旱、低温和热胁迫等胁迫标记。NtGR-RBP1基因在大肠杆菌中表达,随后暴露于胁迫条件下。表达NtGR-RBP1的重组大肠杆菌对盐度、干旱、寒冷和热休克等胁迫更具耐受性。在斑点试验中,重组体与对照相比表现出更高的生长速率。通过监测液体培养试验中的生长情况进一步证实了这种耐受性。在盐(500 mM NaCl)、干旱(20% PEG)、寒冷(4和20°C)和热胁迫(50°C)条件下表达NtGR-RBP1的细胞具有增强的生长能力和更好的耐受性。我们的研究支持这样一种观点,即NtGR-RBP1的保护作用可能有助于在多种环境胁迫下的生长和存活。

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