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Published on: October 20, 2023
[Construction and expression of the eukaryotic expression vector containing AIF gene].
Wen-Wen Deng1, Shu-Kun Liu, Zun-Zhen Zhang
1Department of Environmental Health, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Summary
Researchers successfully constructed an apoptosis-inducing factor (AIF) gene expression vector in A549 cells. This AIF-pcDNA3.1(+) vector enables further investigation into the role of AIF in cellular processes.
Area of Science:
- Molecular Biology
- Gene Expression
- Cancer Research
Background:
- Apoptosis-Inducing Factor (AIF) plays a critical role in programmed cell death.
- Understanding AIF gene function requires effective expression systems in relevant cell lines.
Purpose of the Study:
- To construct a eukaryotic expression vector for the apoptosis-inducing factor (AIF) gene.
- To investigate the expression of the AIF gene in A549 lung carcinoma cells.
Main Methods:
- Amplification of the AIF gene from A549 cells using RT-PCR.
- Cloning the AIF gene into pUC-T and subsequently into the pcDNA3.1(+) eukaryotic expression vector.
- Transfection of the AIF-pcDNA3.1(+) vector into A549 cells and verification of AIF gene expression via RT-PCR and Western blot.
Main Results:
- Successful amplification and cloning of the AIF gene into the AIF-pcDNA3.1(+) vector.
- Sequence analysis confirmed the cloned AIF fragment matched the GenBank sequence.
- AIF gene expression was significantly higher in A549 cells transfected with AIF-pcDNA3.1(+) compared to control cells.
Conclusions:
- The AIF eukaryotic expression vector, AIF-pcDNA3.1(+), was successfully constructed and validated in A549 cells.
- This engineered vector provides a foundation for further research into the biological functions of the AIF gene.
