Related Experiment Video
Updated: Aug 10, 2026
05:52
In Situ Measurement and Correlation of Cell Density and Light Emission of Bioluminescent Bacteria
Published on: June 28, 2018
[Construction of Streptococcus mutans luxS gene allelic exchange plasmid]
1Dept. of Oral Medicine, The Ninth Affiliated Hospital of Shanghai Jiaotong University School of Medicine, Shanghai 200011, China.
Summary
Researchers constructed a Streptococcus luxS gene allelic exchange plasmid for creating knockout mutants. This tool is crucial for studying Streptococcus mutans quorum sensing and interspecies communication.
Area of Science:
- Microbiology
- Molecular Biology
Background:
- The Streptococcus mutans luxS gene is implicated in interspecies quorum sensing.
- Understanding quorum sensing is vital for controlling bacterial communication and virulence.
Purpose of the Study:
- To construct an allelic exchange plasmid for the Streptococcus mutans luxS gene.
- To facilitate the creation of a luxS gene knockout mutant in S. mutans.
Main Methods:
- Utilized nest PCR to amplify upstream and downstream flanking DNA fragments of the luxS gene.
- Employed double endonuclease digestion and ligation to insert fragments into a pBluescript SK (+) Phagemid vector.
- Incorporated the E. coli kanamycin resistance gene for selection.
Main Results:
- Successfully constructed the Xukd-pbsk plasmid containing the luxS flanking regions and kanamycin resistance gene.
- Confirmed plasmid integrity and correct insertion via endonuclease digestion and DNA sequencing.
- Demonstrated in vitro expression of the kanamycin resistance gene.
Conclusions:
- The constructed S. mutans luxS gene allelic exchange plasmid is validated and functional.
- This plasmid serves as a key tool for future research on S. mutans luxS gene function.
- Enables the generation of S. mutans luxS knockout mutants for further study.
