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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
DNA as a Genetic Template02:05

DNA as a Genetic Template

Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
DNA as a Genetic Template02:05

DNA as a Genetic Template

Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...

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One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster
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Published on: January 26, 2024

T-DNA as a gene tag.

Walden Richard1, Hiroaki Hayashi1, Jeff Schell1

  • 1Max-Planck-institut für Züchtungsforschung, Carl von Linne Weg 10, D5000 Kö ln 30, Germany.

The Plant Journal : for Cell and Molecular Biology
|July 9, 2011
PubMed
Summary

Agrobacterium-mediated plant transformation utilizes T-DNA as an insertional mutagen for gene tagging. This method enables the development of transgenic plants with specific, selectable mutations for genetic research.

Area of Science:

  • Molecular Biology
  • Genetics
  • Plant Science

Background:

  • Agrobacterium-mediated plant transformation is a key technique in plant genetics.
  • Transfer DNA (T-DNA) from Agrobacterium tumefaciens can integrate into the plant genome.
  • T-DNA insertion can disrupt gene function, creating mutants.

Purpose of the Study:

  • To explore the utility of T-DNA as an insertional mutagen and gene tag.
  • To investigate the potential of T-DNA tagging for isolating specific mutations.
  • To highlight the application of T-DNA vectors in generating selectable mutations in transgenic plants.

Main Methods:

  • Utilizing Agrobacterium-mediated transformation.
  • Employing T-DNA as an insertional mutagen.

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  • Developing T-DNA-based vectors for gene tagging.

    • Main Results:

    • T-DNA insertion has successfully generated a growing number of plant mutants.
    • The experimental system demonstrates versatility in tagging DNA sequences that regulate gene expression.
    • T-DNA tagging offers potential for isolating specific mutations, moving beyond fortuitous discovery.

    Conclusions:

    • T-DNA-mediated plant transformation is a powerful tool for insertional mutagenesis and gene discovery.
    • This approach facilitates the identification and isolation of genes controlling gene expression.
    • T-DNA-based vectors provide a promising strategy for generating targeted, selectable mutations in plants.