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肺炎克雷伯菌中负责L-山梨糖转运与代谢的sor基因的克隆及物理图谱构建

Cloning and physical mapping of the sor genes for L-sorbose transport and metabolism from Klebsiella pneumoniae.

作者信息

Wöhrl B M, Lengeler J W

机构信息

Max-Planck-Institut für Molekulare Genetik, Berlin, FRG.

出版信息

Mol Microbiol. 1990 Sep;4(9):1557-65. doi: 10.1111/j.1365-2958.1990.tb02067.x.

Abstract

The sor genes of Klebsiella pneumoniae KAY2026, which enable the bacterium to metabolize the ketose L-sorbose, have been cloned on an 8.3kb DNA fragment into the multicopy plasmid, pACYC184. The genes were mapped by restriction analysis, by deletion mapping and by insertion mutagenesis with Tn1725. The corresponding gene products were identified by the maxicell technique. The structural genes sorD, sorA and sorE code for a D-glucitol-6-P dehydrogenase (27 kilodalton (kD)), an Enzymell (EllSor) activity specific for L-sorbose and an L-sorbose-1-P reductase (45kD). Besides these genes for known functions, three additional genes were discovered: sorC, coding for a transcriptional 40kD regulatory protein, and sorF and sorB, coding for two proteins of 14kD and 19kD, respectively, involved in transport. The genes form an operon (gene order sorCpCDFBAE) and are inducible by L-sorbose.

摘要

肺炎克雷伯菌KAY2026的sor基因可使该细菌代谢酮糖L-山梨糖,这些基因已被克隆到一个8.3kb的DNA片段上,并插入多拷贝质粒pACYC184中。通过限制性分析、缺失作图以及用Tn1725进行插入诱变对这些基因进行了定位。通过大细胞技术鉴定了相应的基因产物。结构基因sorD、sorA和sorE分别编码D-葡萄糖醇-6-磷酸脱氢酶(27千道尔顿(kD))、对L-山梨糖具有特异性的酶II(EllSor)活性以及L-山梨糖-1-磷酸还原酶(45kD)。除了这些具有已知功能的基因外,还发现了另外三个基因:sorC,编码一种40kD的转录调节蛋白;sorF和sorB,分别编码参与转运的14kD和19kD的两种蛋白质。这些基因形成一个操纵子(基因顺序为sorCpCDFBAE),并可被L-山梨糖诱导。

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