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IPTG, Dioxane-Free

Company: Promega
Catalog#: V3951
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Stationary-phase Mutagenesis Soft-agar Overlay Assays in Bacillus subtilis
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2017-12-05
[Abstract]  Elucidating how a population of non-growing bacteria generates mutations improves our understanding of phenomena like antibiotic resistance, bacterial pathogenesis, genetic diversity and evolution. To evaluate mutations that occur in nutritionally stressed non-growing bacteria, we have employed the strain B. subtilis YB955, which measures the reversions rates to the chromosomal auxotrophies hisC952, metB5 and leuC427 (Sung and Yasbin, 2002). This gain-of-function system has successfully allowed establishing the role played by repair systems and transcriptional factors in stress-associated mutagenesis (SPM) (Barajas-Ornelas et al., 2014; Gómez-Marroquín et al., 2016). In a recent study (Castro-Cerritos et al., 2017), it was ... [摘要]  阐明非增长细菌群体如何产生突变提高了我们对诸如抗生素抗性,细菌发病机理,遗传多样性和进化等现象的理解。为了评估在营养强调的非生长细菌中发生的突变,我们使用了菌株B。它们测量对染色体营养缺陷型hisC952,metB5和leuC427(Sung和Yasbin,2002)的逆转速率。这种功能获得性系统已成功地允许建立修复系统和转录因子在压力相关的诱变(SPM)中所起的作用(Barajas-Ornelas等人,2014;Gómez-Marroquín等。,2016)。在最近的研究(Castro-Cerritos等人,2017)中,发现核糖核苷酸还原酶(RNR)是SPM所必需的;这种酶在这种细菌中是必不可少的。我们设计了菌株B的条件性突变体。其中通过异丙基-β-D-硫代半乳糖吡喃糖苷(IPTG)(Castro-Cerritos等人,2017)调控了nrdEF操纵子的表达, 。确定在该条件突变体中在营养胁迫下确定三个基因(hisC952,metB5,leuC427)中的氨基酸原养型的突变频率的条件详细这里。这种技术可以用来评估重要基因在应激B发生的致突变过程中的参与。枯草杆菌细胞。

【背景】涉及DNA复制的大约270个基因,包括dnaA,dnaB,dnaC和nrdEF操纵子,它编码RNR,被认为是必不可少的。枯草芽孢杆菌生长(Kobayashi等人,2003)。在这里我们描述了一个协议,已被用来了解这种酶的作用调节事件诱变事件营养应激的非生长细菌菌株枯草芽孢杆菌YB955(emC952,emCecuC427, ...

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