Topological complexity of different populations of pBR322 as visualized by two-dimensional agarose gel electrophoresis
| Autor: | Luis Martin-Parras, Dora B. Krimer, Jorge Bernardo Schvartzman, Isabelle Lucas, Pablo E. Hernández, Olivier Hyrien, María Luisa Martínez-Robles |
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| Rok vydání: | 1998 |
| Předmět: |
Xenopus
Intertwined catenated Dimers chemistry.chemical_compound Plasmid Genetics Escherichia coli Animals Electrophoresis Gel Two-Dimensional Processive recombination Electrophoresis Agar Gel Sv40 Dna-Replication Xenopus Eggs biology DNA Superhelical Topoisomerase Strand Exchange biology.organism_classification Coli topoisomerase-I Biochemistry chemistry Potential origins Agarose gel electrophoresis biology.protein Agarose Nucleic Acid Conformation Ethidium bromide Deoxyribonuclease I DNA Plasmids Research Article |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | 9 páginas, 5 figuras, -- PAGS nros. 3424-3432 Neutral/neutral two-dimensional (2D) agarose gel electrophoresis was used to investigate populations of the different topological conformations that pBR322 can adopt in vivo in bacterial cells as well as in Xenopus egg extracts. To help in interpretation and identification of all the different signals, undigested as well as DNA samples pretreated with DNase I, topoisomerase I and topoisomerase II were analyzed. The second dimension of the 2D gel system was run with or without ethidium bromide to account for any possible changes in the migration behavior of DNA molecules caused by intercalation of this planar agent. Finally, DNA samples were isolated from a recA(-) strain of Escherichia coil, as well as after direct labeling of the replication intermediates in extracts of Xenopus laevis eggs. Altogether, the results obtained demonstrated that 2D gels can be readily used to identify most of the complex topological populations that circular molecules can adopt in vivo in both bacteria and eukaryotic cells. Thiswork was partially supported by grants 96/0470 and PM95/0016from the Spanish Fondo de Investigación Sanitaria and DirecciónGeneral de Enseñanza Superior respectively and grants from theFrench CNRS-ATIPE, ARC and LNCC. I.L. was supported by aMENESR fellowship. |
| Databáze: | OpenAIRE |
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