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天蓝色链霉菌抗生素产生所需转录本的水平有条件地依赖于一个tRNA基因。

The level of a transcript required for production of a Streptomyces coelicolor antibiotic is conditionally dependent on a tRNA gene.

作者信息

Guthrie E P, Chater K F

机构信息

John Innes Institute, John Innes Centre for Plant Science Research, Norwich, United Kingdom.

出版信息

J Bacteriol. 1990 Nov;172(11):6189-93. doi: 10.1128/jb.172.11.6189-6193.1990.

Abstract

In Streptomyces coelicolor A3(2), bldA mutants are conditionally defective in aerial mycelium formation and fail to synthesize all four antibiotics produced by bldA+ strains. Previous studies showed that bldA specifies the tRNA for the rarely used leucine codon UUA. Here we describe experiments examining the abundance in a bldA mutant of a transcript involved in antibiotic production. With use of a bacteriophage-based integrative vector, a promotorless xylE reporter gene was inserted into a previously undescribed gene for an early step in biosynthesis of the red antibiotic undecylprodigiosin, located in the red gene cluster. With this transcriptional fusion present at unit copy number in the chromosome, xylE expression in a bldA+ strain was maximal late in growth in a liquid production medium and was virtually absent in a bldA mutant. On plates of a different medium, the bldA mutant was able to produce undecylprodigiosin and to express the red::xylE fusion, but both abilities were repressed by increasing the concentration of phosphate in the medium. These experiments showed that the undecylprodigiosin deficiency of bldA mutants cannot be accounted for by the presence of TTA codons in the red structural genes, but rather that bldA influences red gene mRNA abundance. In low-phosphate conditions, an alternative regulatory pathway can lead to red gene expression.

摘要

在天蓝色链霉菌A3(2)中,bldA突变体在气生菌丝体形成方面存在条件性缺陷,并且无法合成bldA+菌株产生的所有四种抗生素。先前的研究表明,bldA指定了用于罕见使用的亮氨酸密码子UUA的tRNA。在这里,我们描述了检测参与抗生素生产的转录本在bldA突变体中的丰度的实验。利用基于噬菌体的整合载体,将一个无启动子的xylE报告基因插入到红色基因簇中红色抗生素十一烷基灵菌红素生物合成早期步骤的一个先前未描述的基因中。由于这种转录融合以单拷贝数存在于染色体中,xylE在bldA+菌株中的表达在液体生产培养基中生长后期达到最大值,而在bldA突变体中几乎不存在。在不同培养基的平板上,bldA突变体能够产生十一烷基灵菌红素并表达红色::xylE融合体,但这两种能力都因增加培养基中磷酸盐的浓度而受到抑制。这些实验表明,bldA突变体的十一烷基灵菌红素缺陷不能由红色结构基因中TTA密码子的存在来解释,而是bldA影响红色基因mRNA的丰度。在低磷酸盐条件下,一条替代的调节途径可以导致红色基因的表达。

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