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[龟裂链霉菌对卡那霉素耐药性的决定因素:染色体上的扩增及可逆性遗传不稳定性]

[Determinant of resistance to kanamycin in Streptomycin rimosus: amplification in the chromosome and reversible genetic instability].

作者信息

Potekhin Ia A, Danilenko V N

出版信息

Mol Biol (Mosk). 1985 May-Jun;19(3):805-17.

PMID:2993855
Abstract

The study on the kanamycin resistance determinant (Kanr) in an oxytetracycline--producing strain of S. rimosus showed that it was capable of amplifying in the chromosome during selection for increasing the antibiotic resistance level. The amplification of the DNA fragment with a molecular weight of 10.3 MDa containing Kanr amounted to 300 copies per genome, which resulted in a more than 1000-fold increase in kanamycin resistance level. Cloning of the Kanr determinant on plasmid SLP1.2 in S. lividans strain 66 was performed. In Streptomyces lividans strain 66 the Kanr determinant preserved the capacity for amplification in the hybrid plasmid pSU10 integrated into the chromosome. The Kanr determinant in the strains of S. rimosus and S. lividans was characterized by transfers Kanr in equilibrium Kans with a frequency of 1 X 10(-3). It was shown that the mutation in S. lividans strain 66 resulting in phenotype Kans was not connected with the structural Kanr gene on plasmid pSU10 but was localized on the chromosome. Phenotype Kans was promoted by a decrease in the number of the copies of the regulatory genetic element designated RES1. The reverse to phenotype Kanr might be due to one of the following events: amplification to the initial level of RES1 and amplification up to 200 copies per the genome of the hybrid plasmid pSU10 containing the Kanr determinant. Amplification of the Kanr determinant with preserved initial level of RES1 element resulted in a more than 1000 times increase in the resistance level.

摘要

对龟裂链霉菌土霉素生产菌株中卡那霉素抗性决定簇(Kanr)的研究表明,在选择提高抗生素抗性水平的过程中,它能够在染色体中进行扩增。含有Kanr的分子量为10.3 MDa的DNA片段的扩增达到每个基因组300个拷贝,这导致卡那霉素抗性水平提高了1000倍以上。将Kanr决定簇克隆到变铅青链霉菌66菌株的质粒SLP1.2上。在变铅青链霉菌66菌株中,Kanr决定簇在整合到染色体中的杂交质粒pSU10中保留了扩增能力。龟裂链霉菌和变铅青链霉菌菌株中的Kanr决定簇的特征是,以1×10⁻³的频率将Kanr转移到平衡的Kans中。结果表明,变铅青链霉菌66菌株中导致Kans表型的突变与质粒pSU10上的结构Kanr基因无关,而是定位在染色体上。表型Kans是由指定为RES1的调控遗传元件拷贝数减少所促进的。向Kanr表型逆转可能是由于以下事件之一:RES1扩增到初始水平,以及含有Kanr决定簇的杂交质粒pSU10每个基因组扩增到200个拷贝。在RES1元件保持初始水平的情况下,Kanr决定簇的扩增导致抗性水平提高了1000倍以上。

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