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omp 基因增强了苜蓿中华根瘤菌 CCNWSX0020 对 Cu(II)的细胞耐受性。

An omp gene enhances cell tolerance of Cu(II) in Sinorhizobium meliloti CCNWSX0020.

机构信息

State Key Laboratory of Crop Stress Biology in Arid Areas, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China.

出版信息

World J Microbiol Biotechnol. 2013 Sep;29(9):1655-60. doi: 10.1007/s11274-013-1328-y. Epub 2013 Mar 24.

Abstract

The main aim of this work was to study molecular characterization of a DNA fragment conferring resistance to Cu(II) in Sinorhizobium meliloti CCNWSX0020. The strain CCNWSX0020, resistant to 1.4 mmol l(-1) Cu(II) in tryptone-yeast extract medium was isolated from Medicago lupulina growing in mine tailings of Fengxian County, China. The availability of the complete genome sequence of S. meliloti CCNWSX0020 provides an opportunity for investigating genes that play significant roles in Cu(II) resistance. A copper resistance gene, with a length of 1,445 bp, encoding 481 amino acids, designated omp, was identified by cDNA-amplified fragment length polymorphism from S. meliloti CCNWSX0020. The expression of omp gene strongly increased in the presence of Cu(II). The omp-defective mutants display sensitivities to Cu(II) compared with their wild types. The Cu(II)-sensitive phenotype of the mutant was complemented by a 1.5-kb DNA fragment containing omp gene. BLAST analysis revealed that this gene encoded a hypothetical outer membrane protein with 75 % similarity to outer membrane efflux protein in Rhizobium leguminosarum bv. viciae 3841. These studies suggested that the omp product was involved in the Cu(II) tolerance of S. meliloti CCNWSX0020.

摘要

本工作的主要目的是研究在中华根瘤菌 CCNWSX0020 中赋予铜(II)抗性的 DNA 片段的分子特征。在含有 1.4mmol·l(-1) Cu(II) 的色氨酸-酵母提取物培养基中,对 1.4mmol·l(-1) Cu(II) 具有抗性的 CCNWSX0020 菌株是从中国丰县矿区生长的紫花苜蓿中分离出来的。中华根瘤菌 CCNWSX0020 的完整基因组序列的可用性为研究在铜(II)抗性中起重要作用的基因提供了机会。通过 cDNA 扩增片段长度多态性从 S. meliloti CCNWSX0020 中鉴定了一个长度为 1,445bp、编码 481 个氨基酸的铜抗性基因,命名为 omp。omp 基因的表达在存在 Cu(II)时强烈增加。omp 缺陷突变体与野生型相比对 Cu(II)更敏感。omp 基因缺失突变体的 Cu(II)敏感表型可通过包含 omp 基因的 1.5kb DNA 片段互补。BLAST 分析表明,该基因编码一个假定的外膜蛋白,与 Rhizobium leguminosarum bv. viciae 3841 的外膜外排蛋白具有 75%的相似性。这些研究表明,omp 产物参与了 S. meliloti CCNWSX0020 对 Cu(II)的耐受。

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