Video Experimental Relacionado

Updated: May 5, 2026

Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach

Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach

Published on: December 14, 2019

La multimerización de plasmidos con un alto número de copias causa inestabilidad: CoIE1 codifica un determinante

D K Summers, D J Sherratt

|April 1, 1984

Resumen

Los vectores de clonación de plásmidos derivados de CoIE1 son inestables debido a la multimerización. El plásmido CoIE1

Área de la Ciencia:

Biología Molecular Biología Molecular
Genética La genética.

Sus antecedentes:

El plásmido multicopia natural CoIE1 exhibe una alta estabilidad, a diferencia de los vectores de clonación relacionados que son propensos a la pérdida.
La inestabilidad del plásmido en vectores relacionados está relacionada con la partición aleatoria y el aumento de la multimerización durante la división celular.
Los multiméricos se mantienen en números de copias más bajos que los monómeros, lo que lleva a la segregación de células libres de plásmidos.

Objetivo del estudio:

Para investigar la base molecular de la estabilidad del plásmido CoIE1.
Identificar los determinantes genéticos responsables de prevenir la multimerización del plásmido y garantizar su estabilidad.
Para mejorar la estabilidad de los vectores de clonación de plásmidos relacionados.

Principales métodos:

Análisis de las dinámicas de partición y multimerización de plásmidos.
Localización de los determinantes de estabilidad y monomerización dentro del genoma CoIE1.
Clonado de determinantes de estabilidad identificados en vectores plásmidos para la evaluación funcional.

Principales resultados:

La estabilidad del plásmido está inversamente correlacionada con la multimerización; la reducción de los multiméricos mejora la estabilidad.
CoIE1 codifica un determinante "cer" esencial para la recombinación independiente de recA, recF y recE, convirtiendo eficientemente los multiméricos en monómeros.
Se identificó una región de 0,38 kb que contiene determinantes de monomerización y estabilidad en CoIE1.
La clonación de esta región en vectores plásmidos aumentó significativamente su estabilidad.

Conclusiones:

El determinante "cer" y la región asociada son cruciales para la estabilidad del plásmido CoIE1 al resolver multiméricos.
La ingeniería de vectores de plásmidos con estos determinantes CoIE1 puede superar los problemas de inestabilidad.
Este hallazgo tiene implicaciones para mejorar la confiabilidad de las herramientas biotecnológicas basadas en plásmidos.

Más Videos Relacionados

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Published on: September 2, 2025

Videos de Conceptos Relacionados

Translesion DNA Polymerases

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

Genome Copying Errors

Genome Copying Errors

DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their survival. Therefore, the copying errors are checked and repaired at three levels.

Condensins

Condensins

Condensins are large protein complexes that use ATP to fuel the assembly of chromosomes during mitosis. They transform the tangled, shapeless mass of post-interphase DNA into individualized chromosomes by compacting, organizing, and segregating chromosomal DNA.
The plant and animal cells contain two types of condensin complexes—condensin I and condensin II. Both complexes have five subunits: two SMC (Structural Maintenance of Chromosomes) subunits, a kleisin subunit, and two HEAT-repeat...

Coordination of Gene Expression Processes in Bacteria

Coordination of Gene Expression Processes in Bacteria

The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...

Plasmids

Plasmids

Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...