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Related Concept Videos

Transduction01:16

Transduction

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Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome...
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Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...
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Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
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Related Experiment Video

Updated: Oct 22, 2025

Scalable Transfection of Maize Mesophyll Protoplasts
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[Transfection]

T Ohta1

  • 1Basic Medical Sciences, University of Tsukuba, Japan.

Nihon Rinsho. Japanese Journal of Clinical Medicine
|March 1, 1996
PubMed
Summary
This summary is machine-generated.

Calcium phosphate transfection is a simple method to introduce DNA into mammalian cells for protein expression analysis. This technique was used to study membrane proteins like Na+, K+-ATPase and Ca+-ATPase.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • Transfection is a key technique for introducing exogenous DNA into mammalian cells.
  • Calcium phosphate-mediated transfection is a widely used, cost-effective method.
  • This method facilitates the study of protein function and structure through gene expression.

Purpose of the Study:

  • To present a protocol for high-efficiency calcium phosphate transfection.
  • To demonstrate its application in analyzing membrane proteins, specifically Na+, K+-ATPase and Ca+-ATPase.
  • To elucidate functional domains and interactions within these ion pumps.

Main Methods:

  • High-efficiency calcium phosphate-mediated DNA transfection protocol.
  • Chimeric gene expression analysis.
  • Site-directed mutagenesis.
  • Analysis of protein expression and function in transfected mammalian cells.

Main Results:

  • The N-terminus of the alpha-subunit of Na+, K+-ATPase acts as a sodium sensor.
  • The ouabain binding site in Na+, K+-ATPase is located between residues 69 and 200.
  • Assembly sites for alpha- and beta-subunits of Na+, K+-ATPase are within 26 residues.
  • A novel Ca+-ATPase isoform exhibits similar ATP affinity to known isoforms.
  • The calmodulin-binding site of Ca+-ATPase interacts with the alpha-subunit of Na+, K+-ATPase.

Conclusions:

  • Calcium phosphate transfection is a versatile tool for studying ion pump structure-function relationships.
  • Detailed functional mapping of Na+, K+-ATPase and Ca+-ATPase has been achieved.
  • Interactions between different ion pump subunits and regulatory proteins can be investigated.