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来自聚球藻属PCC 6803菌株的编码葡糖基甘油磷酸合酶的ggpS基因参与渗透调节物质的合成。

The ggpS gene from Synechocystis sp. strain PCC 6803 encoding glucosyl-glycerol-phosphate synthase is involved in osmolyte synthesis.

作者信息

Marin K, Zuther E, Kerstan T, Kunert A, Hagemann M

机构信息

Universität Rostock, FB Biologie, D-18051 Rostock, Germany.

出版信息

J Bacteriol. 1998 Sep;180(18):4843-9. doi: 10.1128/JB.180.18.4843-4849.1998.

Abstract

A salt-sensitive mutant of Synechocystis sp. strain PCC 6803 defective in the synthesis of the compatible solute glucosylglycerol (GG) was used to search for the gene encoding GG-phosphate synthase (GGPS), the key enzyme in GG synthesis. Cloning and sequencing of the mutated region and the corresponding wild-type region revealed that a deletion of about 13 kb occurred in the genome of mutant 11. This deletion affected at least 10 open reading frames, among them regions coding for proteins showing similarities to trehalose (otsA homolog)- and glycerol-3-phosphate-synthesizing enzymes. After construction and characterization of mutants defective in these genes, it became obvious that an otsA homolog (sll1566) (T. Kaneko et al., DNA Res. 3:109-136, 1996) encodes GGPS, since only the mutant affected in sll1566 showed salt sensitivity combined with a complete absence of GG accumulation. Furthermore, the overexpression of sll1566 in Escherichia coli led to the appearance of GGPS activity in the heterologous host. The overexpressed protein did not show the salt dependence that is characteristic for the GGPS in crude protein extracts of Synechocystis.

摘要

使用集胞藻6803株系(Synechocystis sp. strain PCC 6803)的一个对盐敏感的突变体来寻找编码葡糖基甘油(GG)合成关键酶GG-磷酸合酶(GGPS)的基因,该突变体在相容性溶质葡糖基甘油(GG)的合成方面存在缺陷。对突变区域和相应野生型区域进行克隆和测序后发现,突变体11的基因组中发生了约13 kb的缺失。该缺失至少影响了10个开放阅读框,其中包括与海藻糖(otsA同源物)和甘油-3-磷酸合成酶具有相似性的蛋白质编码区域。在构建并鉴定了这些基因缺陷型突变体后,很明显一个otsA同源物(sll1566)(T. Kaneko等人,《DNA研究》3:109 - 136,1996)编码GGPS,因为只有sll1566受影响的突变体表现出盐敏感性且完全没有GG积累。此外,sll1566在大肠杆菌中的过表达导致了异源宿主中出现GGPS活性。过表达的蛋白没有表现出集胞藻粗蛋白提取物中GGPS所特有的盐依赖性。

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