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Related Concept Videos

In vitro Mutagenesis01:16

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
Conservative Site-specific Recombination and Phase Variation02:53

Conservative 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...
Spontaneous and Induced Mutations01:30

Spontaneous and Induced Mutations

Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
Mutagenicity and Carcinogenicity01:25

Mutagenicity and Carcinogenicity

Mutagenicity and carcinogenicity refer to the ability of drugs to cause genetic defects and induce cancer, respectively. The International Agency for Research on Cancer (IARC) classifies agents into four groups based on their carcinogenic potential. Group 1 agents are known human carcinogens; group 2A agents are probably carcinogenic to humans; group 3 agents lack data to support their role in carcinogenesis; and group 4 includes agents for which data support that they are not likely to be...
Mismatch Repair01:20

Mismatch 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...

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Related Experiment Video

Updated: Jun 11, 2026

The Lambda Select cII Mutation Detection System
07:08

The Lambda Select cII Mutation Detection System

Published on: April 26, 2018

ClosTron-targeted mutagenesis.

John T Heap1, Stephen T Cartman, Sarah A Kuehne

  • 1Centre for Biomolecular Sciences, Institute of Infection Immunity and Inflammation, BBSRC Sustainable BioEnergy Centre, School of Molecular Medical Sciences, University of Nottingham, Nottingham, UK.

Methods in Molecular Biology (Clifton, N.J.)
|July 3, 2010
PubMed
Summary
This summary is machine-generated.

Researchers developed the ClosTron, a novel tool for creating genetic mutations in Clostridium bacteria. This method enables rapid, efficient, and stable mutant generation for enhanced functional genomic studies in Clostridium species.

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Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
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Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Published on: February 5, 2019

Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Clostridium species are significant pathogens and possess useful biological functions.
  • Genome sequencing of key Clostridium species is available, but functional genomic studies are limited.
  • Traditional methods for generating clostridial mutants, such as homologous recombination, have been inefficient and yielded unstable results.

Purpose of the Study:

  • To address the limitations in generating clostridial mutants for functional genomic studies.
  • To introduce and evaluate a novel, efficient, and stable method for creating directed mutations in Clostridium species.

Main Methods:

  • Development and application of a recombination-independent strategy utilizing a re-targeted group II intron, specifically the ClosTron system.
  • The ClosTron facilitates positive selection of insertional mutants.

Main Results:

  • The ClosTron system enables the rapid generation of Clostridium mutants in as little as 10 days.
  • Mutant generation using the ClosTron is highly efficient and reproducible.
  • Insertions created by the ClosTron are extremely stable, overcoming limitations of previous methods.

Conclusions:

  • The ClosTron system significantly expands the options for clostridial functional genomic research.
  • This tool overcomes previous hurdles in generating stable and directed clostridial mutants.
  • Facilitates deeper understanding of Clostridium biology and its implications for disease and biotechnology.