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[Construction and expression of eukaryotic expression plasmid pcDNA3.1/hIL-18]
Ling-Hong Kong1, Yao-Chen Li, Yi-Li Wang
1Institute for Cancer Research, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710061,China.
Summary
Researchers successfully constructed the pcDNA3.1/hIL-18 plasmid for expressing human interleukin-18 (hIL-18) in mammalian cells. This achievement provides a foundation for further studies into the anti-tumor effects of hIL-18.
Area of Science:
- Molecular Biology
- Immunology
- Gene Expression
Background:
- Interleukin-18 (IL-18) is a cytokine with significant immunomodulatory functions.
- Understanding IL-18's role in therapeutic applications requires robust expression systems.
- Eukaryotic expression systems are crucial for producing functional cytokines.
Purpose of the Study:
- To construct a novel eukaryotic expression plasmid, pcDNA3.1/hIL-18.
- To achieve successful expression of human interleukin-18 (hIL-18) in mammalian cell lines.
- To lay the groundwork for investigating the anti-tumor potential of IL-18.
Main Methods:
- The hIL-18 cDNA was cloned into the pcDNA3.1 expression vector.
- Liposome-mediated gene transfer was used to transfect COS-7 and Rlc310 cells.
- Immunohistochemical staining and RT-PCR were employed to confirm hIL-18 expression and mRNA levels.
Main Results:
- A recombinant eukaryotic expression plasmid, pcDNA3.1/hIL-18, was successfully constructed.
- Transient expression of hIL-18 was achieved in COS-7 cells.
- Stable expression of hIL-18 was established in Rlc310 cells.
Conclusions:
- The successful construction and expression of the pcDNA3.1/hIL-18 plasmid have been validated.
- This validated system provides a crucial tool for future research.
- The study establishes a foundation for exploring the anti-tumor effects of IL-18.