A deoxyviolacein-based transposon insertion vector for pigmented tracer studies

IF 3.9 3区 生物学 Q2 MICROBIOLOGY MicrobiologyOpen Pub Date : 2024-07-10 DOI:10.1002/mbo3.1425
Benjamin R. Dietz, Tyler J. Nelson, Neil E. Olszewski, Brett M. Barney
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Abstract

Pigments provide a simple means to rapidly visually ascertain the quantities or presence of specific microbes in a complex community. The selection of pigment-producing colonies that are simple to differentiate from common colony phenotypes provides a high degree of certainty for the identity of pigment-tagged strains. Successful employment of pigment production is dependent on various intrinsic factors related to proper levels of gene expression and pigment production that are not always easy to predict and vary within each microbe. We have constructed a simple transposon system that incorporates the genes for the production of deoxyviolacein, a pigment produced from intracellular reserves of the amino acid tryptophan, to randomly insert these genes throughout the genome. This tool allows the user to select from many thousands of potential sites throughout a bacterial genome for an ideal location to generate the desired amount of pigment. We have applied this system to a small selection of endophytes and other model bacteria to differentiate these strains from complex communities and confirm their presence after several weeks in natural environments. We provide two examples of applications using the pigments to trace strains following introduction into plant tissues or to produce a reporter strain for extracellular nitrogen compound sensing. We recognize that this tool could have far broader utility in other applications and microbes, and describe the methodology for use by the greater scientific community.

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基于脱氧紫草素的转座子插入载体,用于色素示踪剂研究。
色素提供了一种简单的方法,可以快速直观地确定复杂群落中特定微生物的数量或存在情况。选择易于与普通菌落表型区分的色素产生菌落,可高度确定色素标记菌株的身份。成功利用色素生产取决于与适当的基因表达水平和色素生产有关的各种内在因素,而这些因素并不总是那么容易预测,而且在每种微生物内部也各不相同。我们构建了一个简单的转座子系统,其中包含了生产脱氧紫草素(一种由细胞内氨基酸色氨酸储备产生的色素)的基因,可在整个基因组中随机插入这些基因。这种工具允许用户从细菌基因组中成千上万个潜在位点中选择一个理想的位置来产生所需数量的色素。我们已将这一系统应用于少量内生菌和其他模式细菌,以将这些菌株从复杂的群落中区分出来,并在自然环境中数周后确认它们的存在。我们提供了两个应用实例,利用色素追踪引入植物组织后的菌株,或生产用于胞外氮化合物传感的报告菌株。我们认识到这一工具在其他应用和微生物中可能有更广泛的用途,并介绍了供广大科学界使用的方法。
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来源期刊
MicrobiologyOpen
MicrobiologyOpen MICROBIOLOGY-
CiteScore
8.00
自引率
0.00%
发文量
78
审稿时长
20 weeks
期刊介绍: MicrobiologyOpen is a peer reviewed, fully open access, broad-scope, and interdisciplinary journal delivering rapid decisions and fast publication of microbial science, a field which is undergoing a profound and exciting evolution in this post-genomic era. The journal aims to serve the research community by providing a vehicle for authors wishing to publish quality research in both fundamental and applied microbiology. Our goal is to publish articles that stimulate discussion and debate, as well as add to our knowledge base and further the understanding of microbial interactions and microbial processes. MicrobiologyOpen gives prompt and equal consideration to articles reporting theoretical, experimental, applied, and descriptive work in all aspects of bacteriology, virology, mycology and protistology, including, but not limited to: - agriculture - antimicrobial resistance - astrobiology - biochemistry - biotechnology - cell and molecular biology - clinical microbiology - computational, systems, and synthetic microbiology - environmental science - evolutionary biology, ecology, and systematics - food science and technology - genetics and genomics - geobiology and earth science - host-microbe interactions - infectious diseases - natural products discovery - pharmaceutical and medicinal chemistry - physiology - plant pathology - veterinary microbiology We will consider submissions across unicellular and cell-cluster organisms: prokaryotes (bacteria, archaea) and eukaryotes (fungi, protists, microalgae, lichens), as well as viruses and prions infecting or interacting with microorganisms, plants and animals, including genetic, biochemical, biophysical, bioinformatic and structural analyses. The journal features Original Articles (including full Research articles, Method articles, and Short Communications), Commentaries, Reviews, and Editorials. Original papers must report well-conducted research with conclusions supported by the data presented in the article. We also support confirmatory research and aim to work with authors to meet reviewer expectations. MicrobiologyOpen publishes articles submitted directly to the journal and those referred from other Wiley journals.
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