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[物种名称]中一个突变体的转录组学和表型分析

Transcriptomic and Phenotypic Analysis of a Mutant in .

作者信息

Diallo Mamou, Kint Nicolas, Monot Marc, Collas Florent, Martin-Verstraete Isabelle, van der Oost John, Kengen Servé W M, López-Contreras Ana M

机构信息

Wageningen Food and Biobased Research, Wageningen, Netherlands.

Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands.

出版信息

Front Microbiol. 2020 Sep 15;11:556064. doi: 10.3389/fmicb.2020.556064. eCollection 2020.

Abstract

SpoIIE is a phosphatase involved in the activation of the first sigma factor of the forespore, σ , during sporulation. A Δ mutant of NCIMB 8052, previously generated by CRISPR-Cas9, did not sporulate but still produced granulose and solvents. Microscopy analysis also showed that the cells of the Δ mutant are elongated with the presence of multiple septa. This observation suggests that in , SpoIIE is necessary for the completion of the sporulation process, as seen in and . Moreover, when grown in reactors, the mutant produced higher levels of solvents than the wild type strain. The impact of the inactivation on gene transcription was assessed by comparative transcriptome analysis at three time points (4 h, 11 h and 23 h). Approximately 5% of the genes were differentially expressed in the mutant compared to the wild type strain at all time points. Out of those only 12% were known sporulation genes. As expected, the genes belonging to the regulon of the sporulation specific transcription factors (σ , σ , σ , σ ) were strongly down-regulated in the mutant. Inactivation of also caused differential expression of genes involved in various cell processes at each time point. Moreover, at 23 h, genes involved in butanol formation and tolerance, as well as in cell motility, were up-regulated in the mutant. In contrast, several genes involved in cell wall composition, oxidative stress and amino acid transport were down-regulated. These results indicate an intricate interdependence of sporulation and stationary phase cellular events in .

摘要

SpoIIE是一种磷酸酶,在芽孢形成过程中参与前芽孢第一个sigma因子σF的激活。先前通过CRISPR-Cas9构建的NCIMB 8052的Δ突变体不形成芽孢,但仍能产生颗粒淀粉和溶剂。显微镜分析还表明,Δ突变体的细胞伸长,存在多个隔膜。这一观察结果表明,在该菌中,SpoIIE对于芽孢形成过程的完成是必需的,如在其他菌中所见。此外,当在反应器中生长时,该突变体产生的溶剂水平高于野生型菌株。通过在三个时间点(4小时、11小时和23小时)进行比较转录组分析,评估了该基因失活对基因转录的影响。与野生型菌株相比,在所有时间点,该突变体中约5%的基因表达存在差异。其中只有12%是已知的芽孢形成基因。正如预期的那样,属于芽孢形成特异性转录因子(σF、σE、σG、σK)调控子的基因在突变体中强烈下调。该基因的失活还导致在每个时间点参与各种细胞过程的基因表达存在差异。此外,在23小时时,参与丁醇形成和耐受性以及细胞运动性的基因在突变体中上调。相反,一些参与细胞壁组成、氧化应激和氨基酸转运的基因下调。这些结果表明该菌中芽孢形成和稳定期细胞事件之间存在复杂的相互依存关系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cc2/7522474/9c3f8d06e946/fmicb-11-556064-g001.jpg

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