Related Experiment Video
Updated: Aug 10, 2026

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
Published on: April 1, 2016
Insertional mutagenesis based on illegitimate recombination in Schizosaccharomyces pombe
G Chua1, L Taricani, W Stangle
1Department of Biology, Biosciences Complex, Queen's University, Kingston, Ontario K7L 3N6, Canada.
Abstract:
An efficient insertional mutagenesis system has been developed for Schizosaccharomyces pombe based on linear PCR-generated cassettes containing selectable markers. It depends upon illegitimate recombination for integration into the genome. Various selectable markers of different sizes can be used to obtain sufficiently high transformation and integration frequencies. Based on Southern blotting, a single insertion is found in each strain and integration sites are broadly distributed in the genome. Sequence analysis of the insert junctions frequently reveals small regions of homology (4-10 bp) between the ends of the integrated cassette and the disrupted gene. The system has been used for simple genetic screens of various types and as a promoter trap for in-frame GFP fusions.
Related Concept Videos
In-vitro Mutagenesis
Gene Conversion
Conservative Site-specific Recombination and Phase Variation
The recognition sites for Cre recombinase called LoxP...
Exon Recombination
Exon shuffling follows “splice frame rules.” Each exon has three reading...
In vitro Mutagenesis
Mismatch Repair
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...

