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首页> 外文期刊>Journal of bacteriology >Molecular cloning and characterization of the genes (pbpA and rodA) responsible for the rod shape of Escherichia coli K-12: analysis of gene expression with transposon Tn5 mutagenesis and protein synthesis directed by constructed plasmids.
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Molecular cloning and characterization of the genes (pbpA and rodA) responsible for the rod shape of Escherichia coli K-12: analysis of gene expression with transposon Tn5 mutagenesis and protein synthesis directed by constructed plasmids.

机译：负责大肠杆菌K-12棒状的基因（pbpA和rodA）的分子克隆和表征：通过转座子Tn5诱变分析基因表达，并构建质粒指导蛋白质合成。

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摘要

Two cell shape-determining genes of Escherichia coli K-12, pbpA, the structural gene for penicillin-binding protein 2, and rodA, whose protein is unknown, were subcloned into plasmid vectors from the transducing phage lambda MAd lip24, which carries the lip-leuS region of the E. coli chromosome. Plasmids with restriction enzyme-created deletions or transposon Tn5 insertions were isolated, and studies of genetic complementation of these plasmids with chromosomal mutations were carried out. Thus, a physical and genetic map of the rodA-pbpA region was established. The genes rodA and pbpA lie side by side within a 4.4-kilobase-pair region. The size of the rodA gene has been shown to be between 0.86 and 1.6 kilobase pairs; such DNA would encode a protein with a molecular weight between 32,000 and 59,000. Since Tn5 mutagenesis of the rodA gene did not affect the expression of the pbpA gene and vice versa, the genes rodA and pbpA seem to have independent promoters. Analysis of the proteins synthesized from the constructed plasmids in maxicells revealed that the plasmid carrying the pbpA gene encoded penicillin-binding protein 2 and amplification of the protein occurred. The product of the rodA gene was not identified.
机译：从转导噬菌体λMAd lip24的质粒载体中，将大肠杆菌K-12的两个细胞形状决定基因pbpA（青霉素结合蛋白2的结构基因）和rodA（其蛋白未知）亚克隆到质粒载体中。大肠杆菌染色体的-leuS区。分离具有限制性酶造成的缺失或转座子Tn5插入的质粒，并对这些具有染色体突变的质粒进行遗传互补研究。因此，建立了rodA-pbpA区的物理和遗传图谱。 rodA和pbpA基因并排位于4.4碱基对区域内。 rodA基因的大小已显示在0.86至1.6千碱基对之间。这样的DNA将编码分子量在32,000和59,000之间的蛋白质。由于rodA基因的Tn5诱变不会影响pbpA基因的表达，反之亦然，因此rodA和pbpA基因似乎具有独立的启动子。对由构建的质粒合成的蛋白质的分析显示其携带了pbpA基因，该质粒编码青霉素结合蛋白2，并发生了扩增。没有鉴定出rodA基因的产物。

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