Employing Cloning-Independent Mutagenesis of Parvimonas micra for the Study of Cell Wall Biogenesis.
Employing Cloning-Independent Mutagenesis of Parvimonas micra for the Study of Cell Wall Biogenesis.
复制标题
利用微小单胞菌的克隆独立诱变来研究细胞壁生物发生。
DOI：
10.1007/978-1-0716-3491-2_5
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Merritt,Justin
中科院分区：
文献类型：
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作者：
Higashi,DustinL;Zou,Zhengzhong;Qin,Hua;Kreth,Jens;Merritt,Justin
细胞壁对细菌起着重要的结构作用，与从生长、分化到致病的各种关键过程密切相关。我们对细胞壁生物发生的理解主要来自于相对较少的大量研究的模式生物。因此，这些过程只能在绝大多数原核生物中推断，特别是在一组没有特征的和/或遗传上难以处理的生物体中。最近，我们为微小细小单胞菌开发了第一个易处理的遗传系统，这是一种普遍存在的人类微生物群的革兰氏阳性病原体，涉及多种类型的炎性感染和各种恶性肿瘤。微型细小单胞菌也是第一个，也是目前唯一的，在芽孢杆菌的整个Tissierelliaclas成员中，已经证明了有针对性的基因操作。因此，现在有可能研究Tissierellia成员的细胞壁生物发生机制，这也可能揭示新的方面。微病理生物学。在这里，我们描述了一种不依赖克隆的基因操作OFP的过程。MICRA，包括等位基因置换突变和遗传互补。所描述的技术也同样适用于其他方面的研究。微病理生物学和生理学。
The cell wall plays an important structural role for bacteria and is intimately tied to a variety of critical processes ranging from growth and differentiation to pathogenesis. Our understanding of cell wall biogenesis is primarily derived from a relatively small number of heavily studied model organisms. Consequently, these processes can only be inferred for the vast majority of prokaryotes, especially among groups of uncharacterized and/or genetically intractable organisms. Recently, we developed the first tractable genetic system forParvimonas micra, which is a ubiquitous Gram-positive pathobiont of the human microbiome involved in numerous types of inflammatory infections as well as a variety of malignant tumors.P. micrais also the first, and currently only, member of the entireTissierelliaclass of theBacillotaphylum in which targeted genetic manipulation has been demonstrated. Thus, it is now possible to study cell wall biogenesis mechanisms within a member of theTissierellia, which may also reveal novel aspects ofP. micrapathobiology. Herein, we describe a procedure for cloning-independent genetic manipulation ofP. micra, including allelic replacement mutagenesis and genetic complementation. The described techniques are also similarly applicable for the study of other aspects ofP. micrapathobiology and physiology.