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High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
Published on: December 28, 2015
[Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells]
Kang-lian Tan1, Zhi-jie Li, Jing-hua Liu
1Key Laboratory of Functional Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, China. collinton@126.com
Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA
|December 16, 2005
Summary
Researchers successfully created an expression vector for Human Protein C Homolog 2 (HPC2) in HEK293 cells. This engineered vector allows for high expression of HPC2, paving the way for further functional studies.
Area of Science:
- Molecular Biology
- Cell Biology
- Biotechnology
Background:
- The Human Protein C Homolog 2 (HPC2) is a protein of interest for functional studies.
- Efficient expression systems are crucial for investigating protein function in eukaryotic cells.
Purpose of the Study:
- To construct a eukaryotic expression vector for Human Protein C Homolog 2 (HPC2).
- To achieve high-level expression of HPC2 in human embryonic kidney (HEK293) cells.
Main Methods:
- Subcloning of HPC2 gene into a flag-tagged pcDNA3 vector.
- Transfection of the recombinant plasmid pcDNA3-flag/HPC2 into HEK293 cells using lipofectamine.
- Western blotting analysis of cell lysate to confirm protein expression.
Main Results:
- Successful construction of the eukaryotic expression vector for HPC2 confirmed by restriction enzyme digestion and DNA sequencing.
- High expression levels of the HPC2 fusion protein were observed in transfected HEK293 cells.
Conclusions:
- The developed eukaryotic expression vector is functional and enables high HPC2 expression in HEK293 cells.
- This system provides a vital foundation for subsequent functional investigations of HPC2.
