Dna Uptake

DNA uptake is the process by which a cell acquires DNA from its surrounding environment, enabling genetic information to enter a new cellular context. In bacteria, specialized transport systems can bind extracellular DNA and move it across the cell envelope, where the incoming material may be degraded, maintained, or incorporated into the chromosome through homologous recombination. This process, often associated with natural transformation, can introduce traits such as antibiotic resistance or new metabolic capabilities. Studying DNA uptake helps researchers understand horizontal gene transfer, microbial adaptation, genome evolution, and the development of laboratory methods for introducing genetic material into cells.

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JoVE EoE - Bacterial Growth and Techniques

Competence-Induced DNA Uptake in Staphylococcus aureus

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2025

Source: Cafini, F., et.al. Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus. J. Vis. Exp. (2017)This video demonstrates competence-induced DNA uptake in Staphylococcus aureus through natural transformation. A chloramphenicol-sensitive strain engineered to constitutively express a sigma factor is cultured under nutrient-limiting conditions to enhance competence, then incubated with donor DNA carrying a resistance gene. Successful gene transfer is confirmed by the...

Radiolabeled Amino Acid Uptake Assay to Quantify Cellular Uptake of Amino Acids

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2025

This video demonstrates an in vitro technique for estimating amino acid uptake by bone marrow-derived stromal cells. Na+-dependent amino acid transporters mediate the uptake of radiolabeled amino acids inside the cells, and the cellular uptake is estimated by measuring the radioactivity.

Mitochondrial Calcium Uptake Assay: A Plate Reader-Based Method for Measuring Calcium Uptake in Isolated Mitochondria

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2025

This video describes a plate reader-based assay to measure mitochondrial calcium uptake in isolated mitochondria. The assay analyzes the fluorescence signal of a calcium-sensitive fluorescent dye to study the kinetics of mitochondrial calcium uptake and calcium overload.

Measuring Dopamine Uptake in Mouse Brain Synaptosomes Using Radiolabeled Dopamine

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2025

This video demonstrates the assessment of dopamine uptake in synaptosomes harvested from mouse brains. It outlines steps for preparing synaptosomes for dopamine uptake via dopamine transporters, treating them with varying concentrations of radiolabeled dopamine, isolating the synaptosomes, and measuring radioactivity to quantify dopamine uptake.

Extracellular Vesicle Uptake Assay: A Method to Visualize and Quantify Cellular Uptake of Extracellular Vesicles Using 3D Fluorescence Imaging Technique

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2025

In this video, we demonstrate an assay to visualize and quantify the cellular uptake of extracellular vesicles or EVs. This assay utilizes 3D fluorescent imaging using a confocal microscope and allows for distinguishing internalized EVs from ones adhered to the cell surface.

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