A restriction map of naphthalene catabolic plasmid pWW60-1 and the location of some of its catabolic genes
A restriction map of naphthalene catabolic plasmid pWW60-1 and the location of some of its catabolic genes
复制标题
萘分解代谢质粒pWW60-1的限制性图谱及其一些分解代谢基因的位置
DOI：
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发表时间：
1986
期刊：
影响因子：
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通讯作者：
Peter A. Williams
中科院分区：
文献类型：
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作者：
P. Cane;Peter A. Williams
总结：提供了87 kbp IncP 9萘分解代谢质粒pWW 60 -1的限制性酶切图谱。使用直接Tn 5插入或使用涉及RP 4和pWW 60 -1之间的不稳定共整合体的中间形成的间接方法获得转座子突变体。影响早期酶的表达的途径（萘水杨酸）的插入分离的插入影响水杨酸羟化酶（nahG）或儿茶酚2，3-加氧酶（nahH）的表达。将nahABC克隆到质粒的6.9kbp HindIII片段（HE）或5.7kbp XhoI片段（XD）的连续区域上。将nahGH克隆在XhoI片段（XC）上距nahABC约30 kbp的区域上，但nahH仅在来自两个衍生质粒pWW 60 -1的相应但较小的XhoI片段上表达，在该区域中缺失。nahH的详细限制性酶切图谱显示与TOL质粒的儿茶酚2，3-加氧酶基因的限制性酶切图谱没有相似性，尽管克隆的基因确实与来自两种不同TOL质粒的儿茶酚2，3-加氧酶基因杂交。pWW 60 -1上分解代谢基因的组织表明，如萘分解代谢质粒NAH 7（也称为pIG 7）所述，它们分离为两个操纵子，但它们转录的相对方向与NAH 7不同。
SUMMARY: A restriction map for the 87 kbp IncP9 naphthalene catabolic plasmid pWW60-1 is presented. Transposon mutants were obtained using direct Tn5 insertion or using an indirect method involving the intermediate formation of unstable cointegrates between RP4 and pWW60-1. Insertions which affected expression of the early enzymes of the pathway (naphthalene to salicylate) were separated from inserts which affected expression of salicylate hydroxylase (nahG) or catechol 2,3-oxygenase (nahH). nahABC were cloned on a contiguous region of the plasmid on either a 6.9 kbp HindIII fragment (HE) or a 5.7 kbp XhoI fragment (XD). nahGH were cloned on a region situated about 30 kbp from nahABC on an XhoI fragment (XC), but nahH was only expressed on the corresponding but smaller XhoI fragments from two derivative plasmids of pWW60-1 with a deletion in that region. The detailed restriction map of nahH shows no similarities with the restriction maps of the genes for catechol 2,3-oxygenases from TOL plasmids, although the cloned gene did hybridize with genes for catechol 2,3-oxygenase from two different TOL plasmids. The organization of the catabolic genes on pWW60-1 suggests a separation into two operons as also described for the naphthalene catabolic plasmid NAH7 (alternatively called pIG7), but the relative directions of their transcription differs from that in NAH7.