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SecMet-FISH:对产生二次代谢物的微生物进行标记,可视化和列举

Yannick Buijs¹, Aileen Ute Geers¹, Iuliana Nita¹

¹Department of Biotechnology and Biomedicine, Technical University of Denmark, 2800 Kongens Lyngby, Denmark.

FEMS microbiology ecology

|March 15, 2024

概括

二次代谢物 FISH (SecMet-FISH) 可以在不进行培养的情况下检测微生物二次代谢物生产者. 这种方法可视化了单细胞中化合物生物合成的遗传能力,推进了生态研究.

科学领域:

微生物学微生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

培养的局限性阻碍了对微生物二次代谢物的理解.
培养独立的方法对于研究微生物群落至关重要.
二次代谢产物在微生物相互作用和生态系统中起着至关重要的作用.

研究的目的:

开发一种用于检测和量化微生物二次代谢物生产者的新方法.
为了能够对二次代谢物生物合成的遗传能力进行单细胞分析.
克服传统种植方式的局限性.

主要方法:

开发了二次代谢物FISH (SecMet-FISH),将基因向的FISH与溶液混合化相结合.
使用的退化原始向生物合成基因集群 (BGCs) 中的维护腺化 (AD) 和合成酶 (KS) 域.
针对格拉姆阴性和格拉姆阳性细菌进行了优化协议,使流细胞计分析成为可能.

主要成果:

在*Pseuodoalteromonas rubra*细胞中成功可视化了BGCs,证明了SecMet-FISH的可行性.
调整并优化了该协议,使其在细菌细胞悬浮剂中广泛适用.
在一个由五个成员组成的合成社区中,从非生产者中区分出杰出的二次代谢物生产者.

结论:

SecMet-FISH是一个强大的培养独立工具,用于识别微生物二次代谢物生产者.
该方法允许高通量,单细胞定量化生物合成潜力.
SecMet-FISH促进了对微生物群落的研究和自然产品的发现.

关键词:

生物合成基因集群生物合成基因集群光在现场混合化光.功能性基因是指功能性基因.基因鱼 (GeneFISH) 是一种基因鱼.在溶液中的 FISH FISH.二次代谢产物二次代谢产物

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相关概念视频

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...