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In vitro Mutagenesis

In vitro Mutagenesis

In-vitro Mutagenesis

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.

Spontaneous and Induced Mutations

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).

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Related Experiment Video

Updated: Apr 30, 2026

Following the Dynamics of Structural Variants in Experimentally Evolved Populations

Following the Dynamics of Structural Variants in Experimentally Evolved Populations

Published on: February 3, 2023

Signature-tagged mutagenesis.

Irena Kukavica-Ibrulj¹, Roger C Levesque

¹Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, 1030 av. de la médecine, Québec, QC, Canada, G1V 0A6.

Methods in Molecular Biology (Clifton, N.J.)

|May 14, 2014

Summary

Signature-tagged mutagenesis (STM) is a powerful screening tool for identifying bacterial genes. This study adapted STM to find Pseudomonas aeruginosa genes crucial for infection and virulence in vivo.

Area of Science:

Microbiology
Genetics
Infectious Diseases

Background:

Signature-tagged mutagenesis (STM) is a genome-wide functional screening assay.
It identifies mutants with specific phenotypes from mixed populations.
STM can identify genes impacting host adaptation, either diminishing or enhancing it.

Purpose of the Study:

To adapt and apply STM for genome-wide screening of Pseudomonas aeruginosa.
To identify genes essential for the infection process.
To identify genes that enhance Pseudomonas aeruginosa virulence in vivo.

Main Methods:

Utilized a modified Signature-tagged mutagenesis (STM) technique.
Performed genome-wide screening of Pseudomonas aeruginosa.
Applied insertional mutagenesis to identify gene functions.

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Main Results:

Identified genes essential for Pseudomonas aeruginosa infection.
Discovered genes that contribute to the virulence of Pseudomonas aeruginosa in vivo.
Provided a comprehensive genetic landscape of Pseudomonas aeruginosa virulence.

Conclusions:

The modified STM approach is effective for identifying virulence factors in Pseudomonas aeruginosa.
This research enhances understanding of Pseudomonas aeruginosa pathogenesis.
The identified genes are potential targets for therapeutic interventions.