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Analysis of FOXO3a Gene Polymorphism Associated with Asthma
1Department of Biotechnology, Sinhgad College of Engineering, Affiliated to Savitribai Phule Pune University (SPPU), Pune, Maharashtra, India.
Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2018
Summary
This study details a simple Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method for detecting genetic variations in the FOXO3a gene. This cost-effective technique aids in understanding genetic links to inflammatory diseases.
Area of Science:
- Genetics
- Molecular Biology
- Immunology
Background:
- Single-nucleotide polymorphisms (SNPs) are key sources of human genetic variation.
- Identifying disease-associated SNPs aids in diagnostics and predicting drug responses.
- FOXO3a gene is crucial for immunoregulation and linked to inflammatory diseases.
Purpose of the Study:
- To develop and report a Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) protocol.
- To detect a specific polymorphism (rs13217795) in the FOXO3a gene.
- To establish a reliable and cost-effective genotyping method.
Main Methods:
- Genomic DNA amplification of a 667 bp fragment from the FOXO3a gene using PCR.
- Digestion of the amplified fragment with the restriction enzyme PagI.
- Separation of digested fragments via gel electrophoresis to analyze Restriction Fragment Length Polymorphism (RFLP) patterns.
Main Results:
- A specific PCR-RFLP protocol was successfully developed for the FOXO3a gene polymorphism (rs13217795).
- The method allows for the detection of variations by analyzing distinct RFLP patterns after PagI digestion.
- The amplified fragment size was confirmed to be 667 bp.
Conclusions:
- The developed PCR-RFLP protocol is a simple, reliable, and cost-effective method for genotyping FOXO3a gene polymorphisms.
- This method can be valuable for research on the association of FOXO3a variations with immunoregulation and inflammatory diseases.
- The protocol provides a foundational technique for further genetic studies involving this gene.
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