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Screening Foodstuffs for Class 1 Integrons and Gene Cassettes
Published on: June 19, 2015
A novel and rapid method for determining integration frequency catalyzed by integron integrase intI1
Zehua Yang1, Xiaofei Jiang, Quhao Wei
1Center of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
Journal of Microbiological Methods
|September 30, 2008
Summary
Researchers created a faster real-time PCR method to measure gene cassette integration. This technique quantified the high integration frequency mediated by integron integrase intI1, significantly exceeding background levels.
Area of Science:
- Molecular Biology
- Microbiology
- Genetics
Background:
- Integrons are mobile genetic elements that play a crucial role in the dissemination of antibiotic resistance genes.
- Quantifying the frequency of gene cassette integration is essential for understanding integron dynamics and evolution.
- Existing methods for determining integration frequency can be time-consuming and complex.
Purpose of the Study:
- To develop a rapid and user-friendly method for assessing gene cassette integration frequency.
- To accurately quantify the role of integron integrase IntI1 in mediating gene cassette insertion.
- To establish a baseline for background integration events.
Main Methods:
- Development of a real-time fluorescent quantitative PCR (qPCR) assay.
- Utilizing qPCR to measure the integration frequency of the aadA2 gene cassette.
- Comparison of integration frequencies in the presence and absence of integron integrase IntI1.
Main Results:
- The developed real-time qPCR method provides a faster and more convenient approach for determining integration frequency.
- The integration frequency of the aadA2 gene cassette was determined to be 1.87x10(-4) in the presence of integrase IntI1.
- A significantly lower background integration frequency, less than 5.23x10(-8), was observed without the presence of integrase IntI1.
Conclusions:
- The novel real-time qPCR assay is effective for quantifying integron-mediated gene cassette integration.
- Integron integrase IntI1 significantly enhances the frequency of gene cassette integration.
- This method provides a valuable tool for studying the molecular mechanisms of integron function and gene transfer.
