GENETIC-ANALYSIS OF THE ERWINIA-CHRYSANTHEMI-3937 CHRYSOBACTIN IRON-TRANSPORT SYSTEM - CHARACTERIZATION OF A GENE-CLUSTER INVOLVED IN UPTAKE AND BIOSYNTHETIC PATHWAYS
GENETIC-ANALYSIS OF THE ERWINIA-CHRYSANTHEMI-3937 CHRYSOBACTIN IRON-TRANSPORT SYSTEM - CHARACTERIZATION OF A GENE-CLUSTER INVOLVED IN UPTAKE AND BIOSYNTHETIC PATHWAYS
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DOI：
10.1111/j.1365-2958.1991.tb00778.x
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发表时间：
1991-06-01
影响因子：
3.6
通讯作者：
EXPERT, D
中科院分区：
文献类型：
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作者：
FRANZA, T;ENARD, C;EXPERT, D
损害菊欧文氏菌 3937 的金杆菌素铁同化系统的 22 个 Mu dII1734 插入中的 20 个定位于 R-prime 质粒 R'4 中包含的 50 kbp 基因组插入（Enard 等，1988）。 使用接合质粒pULB110 (RP4::mini-Mu)和通用转导噬菌体PHI-EC2，我们在染色体连锁图上定位了该铁转运区和两个未连锁的突变。 Chrysobactin 是一种儿茶酚型铁载体，正如我们之前在大肠杆菌的 entA 基因座中观察到的那样，发现源自菊花的 R'4 可以补充大肠杆菌 entB 和 entE 突变。 亚克隆与大肠杆菌CEBAP15操纵子DNA具有同源性的R'4中的2.9kb Eco Ri和4.8kb Bam H1片段。 这些片段用作 DNA/DNA 杂交探针来筛选野生型基因文库，产生重组粘粒 (pEC7)，能够补充破坏欧文氏菌和埃希氏菌属中 2,3-二羟基苯甲酸生物合成途径的突变。以及大肠杆菌entE突变。 对与这些突变相对应的基因组 Mu dll1734 插入进行物理定位，鉴定出一组基因，这些基因局限于生物合成和受体功能所需的约 10 kb 的 DNA 序列。
Twenty of the twenty-two Mu dII1734 insertions impairing the chrysobactin iron-assimilation system of Erwinia chrysanthemi 3937 were localized to a 50 kbp genomic insert contained in the R-prime plasmid, R'4 (Enard et al., 1988). Using the conjugative plasmid pULB110 (RP4::mini-Mu) and the generalized transducing phage PHI-EC2, we located this iron-transport region and the two unlinked mutations on the chromosome linkage map. Chrysobactin is a catechol-type siderophore and, as we have previously observed with the entA locus of Escherichia coli, the E. chrysanthemi-derived R'4 was found to complement E. coli entB and entE mutations. A 2.9 kb Eco Ri and a 4.8 kb Bam Hl fragment in the R'4 sharing homology with the E. coli entCEBAP15 operon DNA were subcloned. These fragments were used as DNA/DNA hybridization probes to screen a wild-type gene library, yielding a recombinant cosmid (pEC7) able to complement mutations disrupting the 2,3-dihydroxybenzoic acid biosynthetic pathway in both Erwinia and Escherichia spp. as well as the E. coli entE mutation. Physical mapping of the genomic Mu dll1734 insertions corresponding to these mutations led to the identification of a cluster of genes confined to a DNA sequence of about 10 kb required for both biosynthetic and receptor functions.