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突变棉普通应激蛋白-2（GUSP-2）基因赋予大肠杆菌 BL-21 和 CIM-496-棉的多种抗逆性。

Mutant Gossypium universal stress protein-2 (GUSP-2) gene confers resistance to various abiotic stresses in E. coli BL-21 and CIM-496-Gossypium hirsutum.

机构信息

Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Lahore, Pakistan.

Drug Research Program, Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland.

出版信息

Sci Rep. 2021 Oct 14;11(1):20466. doi: 10.1038/s41598-021-99900-x.

DOI:10.1038/s41598-021-99900-x
PMID:34650178
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8516947/
Abstract

Gossypium arboreum is considered a rich source of stress-responsive genes and the EST database revealed that most of its genes are uncharacterized. The full-length Gossypium universal stress protein-2 (GUSP-2) gene (510 bp) was cloned in E. coli and Gossypium hirsutum, characterized and point mutated at three positions, 352-354, Lysine to proline (M1-usp-2) & 214-216, aspartic acid to serine (M2-usp-2) & 145-147, Lysine to Threonine (M3-usp-2) to study its role in abiotic stress tolerance. It was found that heterologous expression of one mutant (M1-usp-2) provided enhanced tolerance against salt and osmotic stresses, recombinant cells have higher growth up to 10-5dilution in spot assay as compared to cells expressing W-usp-2 (wild type GUSP-2), M2-usp-2 and M3-usp-2 genes. M1-usp-2 gene transcript profiling exhibited significant expression (8.7 fold) in CIM-496-Gossypium hirsutum transgenic plants and enhance drought tolerance. However, little tolerance against heat and cold stresses in bacterial cells was observed. The results from our study concluded that the activity of GUSP-2 was enhanced in M1-usp-2 but wipe out in M2-usp-2 and M3-usp-2 response remained almost parallel to W-usp-2. Further, it was predicted through in silico analysis that M1-usp-2, W-usp-2 and M3-usp-2 may be directly involved in stress tolerance or function as a signaling molecule to activate the stress adaptive mechanism. However, further investigation will be required to ascertain its role in the adaptive mechanism of stress tolerance.

摘要

陆地棉是一种富含应激响应基因的资源，EST 数据库显示其大多数基因尚未得到鉴定。我们在大肠杆菌和陆地棉中克隆了全长棉花普遍应激蛋白-2（GUSP-2）基因（510bp），对其进行了鉴定和 3 个位置（352-354、赖氨酸突变为脯氨酸（M1-usp-2）和 214-216、天冬氨酸突变为丝氨酸（M2-usp-2）和 145-147、赖氨酸突变为苏氨酸（M3-usp-2））的定点突变，以研究其在非生物胁迫耐受中的作用。结果发现，一个突变体（M1-usp-2）的异源表达提供了对盐和渗透胁迫的增强耐受，与表达 W-usp-2（野生型 GUSP-2）、M2-usp-2 和 M3-usp-2 基因的细胞相比，重组细胞在点样试验中最高可稀释 10-5 倍生长。M1-usp-2 基因的转录谱分析显示，在 CIM-496-陆地棉转基因植物中显著表达（8.7 倍），并增强了耐旱性。然而，在细菌细胞中观察到对热和冷胁迫的耐受性很小。我们的研究结果表明，M1-usp-2 中的 GUSP-2 活性增强，但在 M2-usp-2 和 M3-usp-2 中丧失，而 W-usp-2 的反应几乎保持平行。此外，通过计算机分析预测，M1-usp-2、W-usp-2 和 M3-usp-2 可能直接参与胁迫耐受或作为信号分子激活应激适应机制。然而，需要进一步的研究来确定其在胁迫耐受适应机制中的作用。

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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f210/8516947/187a48af2918/41598_2021_99900_Fig8_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f210/8516947/e36bd83fd13a/41598_2021_99900_Fig10_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f210/8516947/ee955d9b5038/41598_2021_99900_Fig11_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f210/8516947/c3c3edf868cf/41598_2021_99900_Fig12_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f210/8516947/0c806a611ec4/41598_2021_99900_Fig13_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f210/8516947/d5bc58a86af7/41598_2021_99900_Fig14_HTML.jpg
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