Related Experiment Video
Updated: Aug 1, 2025

Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR
Published on: November 23, 2012
Allelic Exchange: Construction of an Unmarked In-Frame Deletion in Staphylococcus aureus
Merve S Zeden1, Christopher F Schuster2, Angelika Gründling3
1Microbiology, School of Biological and Chemical Sciences, University of Galway, Galway H91 TK33, Ireland.
Abstract:
Here we describe an allelic-exchange procedure for the construction of an unmarked gene deletion in the bacterium Staphylococcus aureus As a practical example, we outline the construction of a tagO gene deletion in S. aureus using the allelic-exchange plasmid pIMAY*. We first present the general principles of the allelic-exchange method, along with information on counterselectable markers. Furthermore, we summarize relevant cloning procedures, such as the splicing by overhang extension (SOE) polymerase chain reaction (PCR) and Gibson assembly methods, and we conclude by giving some general consideration to performing genetic modifications in S. aureus.
Related Concept Videos
Gene Conversion
Conservative Site-specific Recombination and Phase Variation
The recognition sites for Cre recombinase called LoxP...
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...

