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Updated: May 8, 2026
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 of F-box protein 6 gene and its expression in HEK293T cells]
Yuan Gao1, Jinghua Feng, Gan Chen
1Institute of Genetic Engineering, Southern Medical University, Guangzhou, China. gaoyuan1623@163.com
Objective:
To construct eukaryotic expression vector of F-box protein 6 (FBXO6) gene.
Methods:
The full-length FBXO6 cDNA containing two restriction sites (BglII and EcoRI) was synthesized by PCR. The pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 vectors were constructed respectively. The size and sequence of cDNA fragments were confirmed by bacterial colony PCR and BglII and EcoRI digestion and sequencing. HEK293T cells were transfected with the pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 vectors respectively, and then the expression of FBXO6 protein was detected using Western blotting.
Results:
The pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 contained FBXO6 fragments of proper size and sequence. FBXO6 protein was expressed efficiently in pEGFP-C1-FBXO6 transfected HEK293T cells, but it was down-regulated in pEGFP-C1-anti-FBXO6 transfected 293T cells.
Conclusion:
Both pEGFP-C1-FBXO6 and pEGFP-C1-anti-FBXO6 were successfully constructed.
