Overcoming the Protective Barriers of Cells in the Transformation of Bacteria with Exogenous DNA Using the Example of Rhodobacter capsulatus

IF 0.8 Q3 Engineering Nanotechnologies in Russia Pub Date : 2023-09-19 DOI:10.1134/S2635167623700362

E. V. Mayorova, A. I. Romanova, E. P. Petushkova, A. A. Tsygankov

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Abstract

The culturing conditions for the production of cells of the purple nonsulfur bacteria Rba. capsulatus, which possess a minimal capsule, are selected and the effectiveness of their use in induced transformation by electroporation to transfer exogenous DNA in comparison with cells cultivated on other types of conventionally used media is shown. It is found that, upon the electroporation of plasmid DNA (pDNA) into the cells of Rba. capsulatus according to the selected procedure, the survival rate of the culture is an unstable quantity, which requires optimization of the conditions for obtaining electrocompetent cells and careful selection of the electric-shock treatment mode. For the first time, the fundamental possibility of using an approach for protection against the destruction of pDNA by the restriction modification system (RMS) of Rba. capsulatus by means of in vitro methylation with a cell-free extract of this culture, as well as transformation of the treated pDNA into competent cells of this bacterium by electroporation is demonstrated.

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克服外源DNA转化细菌过程中细胞的保护屏障——以荚膜红杆菌为例

紫色无硫细菌Rba细胞的培养条件。选择具有最小胶囊的荚膜，并将其用于电穿孔诱导转化以转移外源DNA的有效性与在其他类型常规使用的培养基上培养的细胞进行比较。发现质粒DNA (pDNA)电穿孔后进入Rba细胞。根据选择的程序，培养物的存活率是一个不稳定的量，这需要优化获得电致能细胞的条件和仔细选择电击处理方式。首次为利用Rba限制性修饰系统(RMS)保护pDNA免受其破坏提供了基本的可能性。通过体外甲基化这种培养的无细胞提取物，以及通过电穿孔将处理过的pDNA转化为这种细菌的能态细胞的方法，证明了荚膜菌。

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来源期刊

Nanotechnologies in Russia NANOSCIENCE & NANOTECHNOLOGY-

CiteScore

1.20

自引率

0.00%

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期刊介绍： Nanobiotechnology Reports publishes interdisciplinary research articles on fundamental aspects of the structure and properties of nanoscale objects and nanomaterials, polymeric and bioorganic molecules, and supramolecular and biohybrid complexes, as well as articles that discuss technologies for their preparation and processing, and practical implementation of products, devices, and nature-like systems based on them. The journal publishes original articles and reviews that meet the highest scientific quality standards in the following areas of science and technology studies: self-organizing structures and nanoassemblies; nanostructures, including nanotubes; functional and structural nanomaterials; polymeric, bioorganic, and hybrid nanomaterials; devices and products based on nanomaterials and nanotechnology; nanobiology and genetics, and omics technologies; nanobiomedicine and nanopharmaceutics; nanoelectronics and neuromorphic computing systems; neurocognitive systems and technologies; nanophotonics; natural science methods in a study of cultural heritage items; metrology, standardization, and monitoring in nanotechnology.