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调控胞外多糖琥珀聚糖和半乳葡聚糖生物合成的苜蓿中华根瘤菌mucR基因的分子分析

Molecular analysis of the Rhizobium meliloti mucR gene regulating the biosynthesis of the exopolysaccharides succinoglycan and galactoglucan.

作者信息

Keller M, Roxlau A, Weng W M, Schmidt M, Quandt J, Niehaus K, Jording D, Arnold W, Pühler A

机构信息

Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Federal Republic of Germany.

出版信息

Mol Plant Microbe Interact. 1995 Mar-Apr;8(2):267-77. doi: 10.1094/mpmi-8-0267.

DOI:10.1094/mpmi-8-0267
PMID:7756693
Abstract

The Rhizobium meliloti Tn5 mutant Rm3131, producing galactoglucan (EPS II) instead of succinoglycan (EPS I), was complemented by a 3.6-kb EcoRI-fragment of the Rhizobium meliloti genome. Sequencing of this fragment revealed six open reading frames (ORFs). The ORF found to be affected in the mutant Rm3131 codes for a putative protein of 15.7 kDa and forms a monocistronic transcriptional unit. Further genetic analysis revealed that the gene mutated in Rm3131 is identical to the previously described R. meliloti mucR gene (H. Zhan, S.B. Levery, C. C. Lee, and J.A. Leigh, 1989, Proc. Natl. Acad. Sci. USA 86:3055-3059). By hybridization it was shown that a mucR homologous gene is present in several rhizobacteria. The deduced amino acid sequence of MucR showed nearly 80% identity to the Agrobacterium tumefaciens Ros protein, a negative regulator of vir genes and necessary for succinoglycan production. MucR contains like Ros a putative zinc finger sequence of the C2H2 type. Transcriptional fusions of genes for EPS I and EPS II synthesis, the so-called exo and exp genes, with the marker gene lacZ were used to delineate the role of mucR for exo and exp gene expression. It was found that exp genes are negatively regulated by MucR on the transcriptional level, whereas a posttranscriptional regulation by MucR is assumed for exo genes. Furthermore, mucR is negatively regulating its own transcription.

摘要

苜蓿中华根瘤菌Tn5突变体Rm3131产生的是半乳葡聚糖(胞外多糖II)而非琥珀聚糖(胞外多糖I),它被苜蓿中华根瘤菌基因组的一个3.6 kb的EcoRI片段互补。对该片段进行测序后发现了6个开放阅读框(ORF)。在突变体Rm3131中发现受影响的ORF编码一个推定的15.7 kDa蛋白质,并形成一个单顺反子转录单元。进一步的遗传分析表明,Rm3131中发生突变的基因与先前描述的苜蓿中华根瘤菌mucR基因相同(H. Zhan、S.B. Levery、C.C. Lee和J.A. Leigh,1989年,《美国国家科学院院刊》86:3055 - 3059)。通过杂交表明,几个根瘤菌中存在mucR同源基因。MucR推导的氨基酸序列与根癌土壤杆菌Ros蛋白显示出近80%的同一性,Ros蛋白是vir基因的负调控因子,也是琥珀聚糖产生所必需的。MucR与Ros一样,含有一个推定的C2H2型锌指序列。利用胞外多糖I和胞外多糖II合成基因(即所谓的exo和exp基因)与标记基因lacZ的转录融合来阐明mucR对外源和exp基因表达的作用。发现exp基因在转录水平上受到MucR的负调控,而推测exo基因受到MucR的转录后调控。此外,mucR对其自身转录具有负调控作用。

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