Episomal Plasmid Curing
Episomal Plasmid Curing - Related Videos
10:52
Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids
0 Views •
Cited by 26 •
2015
Induced pluripotent stem cells (iPSCs) represent a source of patient-specific tissues for clinical applications and basic research. Here, we present a detailed protocol to reprogram human peripheral blood mononuclear cells (PBMNCs) obtained from frozen buffy coats into viral-free iPSCs using non-integrating episomal plasmids.
09:45
Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
0 Views •
Cited by 17 •
2017
This protocol describes a detailed method for efficient generation of integration-free iPSCs from human adult peripheral blood cells. With the use of four oriP/EBNA-based episomal vectors to express the reprogramming factors, KLF4, MYC, BCL-XL, or OCT4 and SOX2, thousands of iPSC colonies can be obtained from 1 mL of peripheral blood.
01:25
Accelerated Curing of Concrete
0 Views •
2024
Accelerating concrete curing is achieved by applying heat and additional moisture. This process accelerates the hydration of the cement, resulting in an earlier strength gain in the concrete. Steam curing is a method wherein the concrete products are either transported through a chamber on a conveyor belt or encased in plastic, allowing steam at atmospheric pressure to circulate freely around them. This process begins with a phase of moist curing that typically lasts between 3 to 5 hours, after...
08:16
Plasmid Purification
0 Views •
2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
01:20
Curing of Concrete
0 Views •
2024
The hydration of cement takes place within the water-filled capillary pores. However, environmental elements can disrupt this process by evaporating water from the concrete surfaces. Sealed concrete with a water-cement ratio below 0.5 experiences self-desiccation, leading to water loss. The water loss in concrete is mitigated by curing. This technique involves keeping the concrete saturated to maintain the necessary temperature and moisture conditions, to optimally fill the spaces in the cement...
FAQs
Without antibiotic selection, cells that lose the episome are no longer disadvantaged by its absence, allowing plasmid-free descendants to accumulate during growth. Because daughter cells can progressively inherit fewer plasmid copies when replication or partitioning is not maintained, repeated cultivation under these conditions can enrich for cured cells. This creates a basis for testing which traits depend on the plasmid.
Conditions such as elevated temperature, chemical stress, and plasmid incompatibility act by disturbing plasmid replication or partitioning. Replication disruption limits production of plasmid copies, whereas partitioning disruption interferes with their distribution to daughter cells. These mechanisms provide alternatives to withholding antibiotic selection when researchers need to favor loss of the episome.
A cured derivative provides a comparison with the original parental strain while retaining the bacterial chromosome as the principal shared genetic background. If a phenotype changes after plasmid removal, the difference supports an association with plasmid-encoded functions rather than a general chromosomal property. This comparison is especially useful for investigating resistance, virulence, or toxin-related traits.
Researchers first grow the bacterial population without antibiotic selection or expose it to a selected curing condition. They then examine cells that have progressively lost the episome and compare the resulting cured population with the parental strain. This paired comparison helps separate plasmid-associated phenotypes from functions retained by the bacterial chromosome.
The resulting cured cells can be compared with parental bacteria for changes in antimicrobial resistance, virulence, and toxin production. Researchers can also assess whether plasmid loss alters interactions between bacteria and their hosts. These comparisons connect the presence of the episome with observable biological traits and help identify phenotypes that depend on mobile genetic elements.
In infection models, comparing cured and parental strains helps test whether a plasmid contributes to bacterial survival, immune evasion, or disease severity. Removing the episome creates an experimental contrast that can link a mobile genetic element to host-pathogen behavior. The approach therefore adds genetic causality to studies of bacterial traits that influence immune responses and infection outcomes.