Related Experiment Video
Updated: Jun 1, 2026
09:35
Analysis of Somatic Hypermutation in the JH4 intron of Germinal Center B cells from Mouse Peyer's Patches
Published on: April 20, 2021
Understanding the immunoglobulin locus specificity of hypermutation
Vera Batrak1, Artem Blagodatski, Jean-Marie Buerstedde
1vera.batrak@googlemail.com
Methods in Molecular Biology (Clifton, N.J.)
|June 11, 2011
Summary
Researchers identified a specific DNA sequence, Diversification Activator (DIVAC), that directs immunoglobulin (Ig) gene hypermutation in B cells. This discovery aids understanding of antibody diversity and immune system function.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Immunoglobulin (Ig) genes undergo high-rate point mutations for antibody affinity maturation, a process crucial for adaptive immunity in jawed vertebrates.
- The precise targeting mechanism of Ig hypermutation to specific gene loci remains a long-standing question in immunology.
- Understanding Ig hypermutation is key to explaining antibody diversity and immune system development.
Purpose of the Study:
- To investigate the locus specificity of Ig hypermutation using a novel experimental system.
- To identify the DNA sequences responsible for directing hypermutation activity to Ig loci.
- To provide a new platform for studying the molecular mechanisms of Ig hypermutation.
Main Methods:
- Development of a chicken B-cell line (DT40) system utilizing a green fluorescent protein (GFP) reporter gene.
- Quantification of somatic hypermutation by measuring loss of GFP fluorescence using fluorescence-activated cell sorting (FACS).
- Targeted integration of the GFP reporter into a defined chromosomal position within the Ig light chain (IgL) locus.
Main Results:
- Identification of a 10 kb sequence within the IgL locus that is necessary and sufficient to activate hypermutation in an adjacent reporter gene.
- This sequence was named Diversification Activator (DIVAC).
- The experimental system demonstrated the ability to quantify hypermutation events and pinpoint locus-specific activators.
Conclusions:
- A novel sequence, DIVAC, has been identified as a key regulator of Ig hypermutation.
- It is postulated that similar cis-acting sequences (DIVAC) are present in Ig loci across jawed vertebrates.
- The developed experimental system offers a promising avenue for further dissecting the molecular mechanisms of Ig hypermutation and identifying functional motifs and trans-acting factors.
More Related Videos
Related Concept Videos
01:31Hybridoma Technology
Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...
01:28Diversity of Antigen Receptors
Antigen receptors are essential components of the immune system crucial in defending the body against foreign invaders. These receptors are present on the surface of B and T cells, enabling them to recognize antigens and mount an appropriate immune response.
Before encountering any antigen, lymphocytes express these receptors. On B cells, the antigen receptor is a membrane-bound antibody molecule called BCR; on T cells, it is a T cell receptor or TCR. B and T cell receptors are composed of two...
Before encountering any antigen, lymphocytes express these receptors. On B cells, the antigen receptor is a membrane-bound antibody molecule called BCR; on T cells, it is a T cell receptor or TCR. B and T cell receptors are composed of two...
01:10Antibody Structure
Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
01:10Antibody Structure
Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
01:20Mismatch Repair
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
02:53Conservative 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...
The recognition sites for Cre recombinase called LoxP...