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过表达嗜冷菌 KOPRI 22215 的冷休克蛋白 A 赋予其抗寒能力。

Overexpression of cold shock protein A of Psychromonas arctica KOPRI 22215 confers cold-resistance.

机构信息

Department of Molecular Biology, Sejong University, 98 Gunja-dong, Kwangjin-gu, Seoul, 143-747, Korea.

出版信息

Protein J. 2010 Feb;29(2):136-42. doi: 10.1007/s10930-010-9233-9.

Abstract

A polar bacterium was isolated from Arctic sea sediments and identified as Psychromonas artica, based on 16S rDNA sequence. Psychromonas artica KOPRI 22215 has an optimal growth temperature of 10 degrees C and a maximum growth temperature of 25 degrees C, suggesting this bacterium is a psychrophile. Cold shock proteins (Csps) are induced upon temperature downshift by more than 10 degrees C. Functional studies have researched mostly Csps of a mesophilic bacterium Escherichia coli, but not on those of psychrophilic bacteria. In an effort to understand the molecular mechanisms of psychrophilic bacteria that allow it withstand freezing environments, we cloned a gene encoding a cold shock protein from P. artica KOPRI 22215 (CspA(Pa)) using the conserved sequences in csp genes. The 204 bp-long ORF encoded a protein of 68 amino acids, sharing 56% homology to previously reported E. coli CspA protein. When CspA(Pa) was overexpressed in E. coli, it caused cell growth-retardation and morphological elongation. Interestingly, overexpression of CspA(Pa) drastically increased the host's cold-resistance by more than ten times, suggesting the protein aids survival in polar environments.

摘要

从北极海洋沉积物中分离到一种极地细菌,并根据 16S rDNA 序列将其鉴定为 Psychromonas artica。Psychromonas artica KOPRI 22215 的最适生长温度为 10°C,最大生长温度为 25°C,表明该细菌是一种嗜冷菌。当温度下降超过 10°C 时,会诱导产生冷休克蛋白(Csps)。功能研究主要针对嗜温菌大肠杆菌的 Csps,但不是针对嗜冷菌的 Csps。为了了解允许嗜冷菌耐受冷冻环境的分子机制,我们使用 csp 基因中的保守序列,从 P. artica KOPRI 22215 中克隆了一个编码冷休克蛋白的基因(CspA(Pa))。该 204 个碱基对的 ORF 编码了一个由 68 个氨基酸组成的蛋白质,与先前报道的大肠杆菌 CspA 蛋白具有 56%的同源性。当 CspA(Pa) 在大肠杆菌中过表达时,它会导致细胞生长减缓和形态伸长。有趣的是,CspA(Pa) 的过表达使宿主的抗寒性提高了十倍以上,表明该蛋白有助于在极地环境中生存。

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