E. V. Mayorova, A. I. Romanova, E. P. Petushkova, A. A. Tsygankov
{"title":"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","authors":"E. V. Mayorova, A. I. Romanova, E. P. Petushkova, A. A. Tsygankov","doi":"10.1134/S2635167623700362","DOIUrl":null,"url":null,"abstract":"<p>The culturing conditions for the production of cells of the purple nonsulfur bacteria <i>Rba. capsulatus</i>, 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 <i>Rba. capsulatus</i> 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 <i>Rba. capsulatus</i> 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.</p>","PeriodicalId":716,"journal":{"name":"Nanotechnologies in Russia","volume":null,"pages":null},"PeriodicalIF":0.8000,"publicationDate":"2023-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Overcoming the Protective Barriers of Cells in the Transformation of Bacteria with Exogenous DNA Using the Example of Rhodobacter capsulatus\",\"authors\":\"E. V. Mayorova, A. I. Romanova, E. P. Petushkova, A. A. Tsygankov\",\"doi\":\"10.1134/S2635167623700362\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>The culturing conditions for the production of cells of the purple nonsulfur bacteria <i>Rba. capsulatus</i>, 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 <i>Rba. capsulatus</i> 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 <i>Rba. capsulatus</i> 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.</p>\",\"PeriodicalId\":716,\"journal\":{\"name\":\"Nanotechnologies in Russia\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.8000,\"publicationDate\":\"2023-09-19\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Nanotechnologies in Russia\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://link.springer.com/article/10.1134/S2635167623700362\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Engineering\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nanotechnologies in Russia","FirstCategoryId":"1085","ListUrlMain":"https://link.springer.com/article/10.1134/S2635167623700362","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Engineering","Score":null,"Total":0}
Overcoming the Protective Barriers of Cells in the Transformation of Bacteria with Exogenous DNA Using the Example of Rhodobacter capsulatus
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.
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.