دورية أكاديمية
Optimisation of DNA electroporation protocols for different plant-associated bacteria.
| العنوان: | Optimisation of DNA electroporation protocols for different plant-associated bacteria. |
|---|---|
| المؤلفون: | Kim EYS; Nitrogen Fixation Laboratory, Department of Biochemistry and Molecular Biology, Federal University of Paraná (UFPR), Curitiba, Brazil., Maltempi de Souza E; Nitrogen Fixation Laboratory, Department of Biochemistry and Molecular Biology, Federal University of Paraná (UFPR), Curitiba, Brazil., Müller-Santos M; Nitrogen Fixation Laboratory, Department of Biochemistry and Molecular Biology, Federal University of Paraná (UFPR), Curitiba, Brazil. Electronic address: marcelomuller@ufpr.br. |
| المصدر: | Journal of microbiological methods [J Microbiol Methods] 2024 May; Vol. 220, pp. 106912. Date of Electronic Publication: 2024 Mar 06. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Biomedical Country of Publication: Netherlands NLM ID: 8306883 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-8359 (Electronic) Linking ISSN: 01677012 NLM ISO Abbreviation: J Microbiol Methods Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam, The Netherlands : Elsevier Biomedical, c1983- |
| مواضيع طبية MeSH: | Transformation, Bacterial* , DNA*, Plasmids ; Electroporation/methods ; Plants ; Bacteria/genetics |
| مستخلص: | Electroporation is a vital process that facilitates the use of modern recombineering and other high-throughput techniques in a wide array of microorganisms, including non-model bacteria like plant growth-promoting bacteria (PGPB). These microorganisms play a significant role in plant health by colonizing plants and promoting growth through nutrient exchange and hormonal regulation. In this study, we introduce a sequential Design of Experiments (DOE) approach to obtain highly competent cells swiftly and reliably for electroporation. Our method focuses on optimizing the three stages of the electroporation procedure-preparing competent cells, applying the electric pulse field, and recovering transformed cells-separately. We utilized a split-plot fractional design with five factors and a covariate to optimize the first step, response surface methodology (RSM) for the second step, and Plackett-Burman design for two categorical factors and one continuous factor for the final step. Following the experimental sequence with three bacterial models, we achieved efficiencies 10 to 100 times higher, reaching orders of 10 5 to 10 6 CFU/μg of circular plasmid DNA. These results highlight the significant potential for enhancing electroporation protocols for non-model bacteria. Competing Interests: Declaration of competing interest None. (Copyright © 2024 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: DNA transformation; Electroporation; Multifactorial optimisation; Plant growth-promoting bacteria (PGPB) |
| المشرفين على المادة: | 9007-49-2 (DNA) |
| تواريخ الأحداث: | Date Created: 20240307 Date Completed: 20240419 Latest Revision: 20240419 |
| رمز التحديث: | 20240419 |
| DOI: | 10.1016/j.mimet.2024.106912 |
| PMID: | 38452904 |
| قاعدة البيانات: | MEDLINE |
Full Text via ClinicalKey 
Full Text Finder كن أول من يترك تعليقا!