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Updated: May 29, 2026
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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 Trim6 eukaryotic expression vector and its expression in HEK293 cells]
Da-Kang Sun1, Xin-Ye An, Feng-Ai Hu
1Experiment Center of Clinical Medicine, Affiliated Hospital of Binzhou Medical University, Binzhou 256603, China.
Summary
Researchers successfully constructed the pcDNA3.1(+)-Trim6 eukaryotic expression vector. This vector enables the expression of Trim6 protein in HEK293T cells, laying groundwork for future studies.
Area of Science:
- Molecular Biology
- Cell Biology
Background:
- The Trim6 protein plays a role in various cellular processes.
- Understanding Trim6 expression is crucial for further research.
Purpose of the Study:
- To construct a recombinant eukaryotic expression vector for Trim6 (pcDNA3.1(+)-Trim6).
- To verify the successful expression of Trim6 in HEK293T cells in vitro.
Main Methods:
- Total RNA was extracted from HeLa cells.
- Reverse transcription polymerase chain reaction (RT-PCR) was used for amplification.
- The target sequences were cloned into the pcDNA3.1(+) vector.
- Recombinant vector confirmation involved restriction enzyme digestion, PCR, and sequencing.
- HEK293T cells were transfected with the recombinant vector.
- Trim6 expression was detected using Western blot analysis.
Main Results:
- Restriction enzyme digestion, PCR, and sequencing confirmed successful vector construction.
- The pcDNA3.1(+)-Trim6 vector demonstrated successful expression of Trim6 protein in HEK293T cells.
Conclusions:
- The successful construction of the pcDNA3.1(+)-Trim6 expression vector was achieved.
- This validated vector provides a foundation for future investigations into the functional roles of Trim6.
