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Updated: Jun 30, 2026
High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
Published on: December 28, 2015
[Construction of a recombinant eukaryotic vector of human intestinal trefoil factor and its expression in 293-T
1Department of Gastroenterology, Zhongshan Hospital, Xiamen University, Xiamen, China. yapi@xmzsh.com
Objective:
To clone human intestinal trefoil factor (hITF/hTFF3) gene into an eukaryotic expression vector for its expression in eukaryotic cells.
Methods:
The total RNA was extracted from normal human colon mucosa, and transcribed into cDNAs using RT-PCR. hTFF3 gene was amplified by PCR and ligated into pGEMT vector by TA cloning method. After sequencing, the hTFF3 gene was transfered into the eukaryotic expression vector pCMV5-myc. The recombinant vector was transfected into 293-T cells, and the expression of the recombinant protein was detected by Western blotting.
Results And Conclusion:
hTFF3 gene was successfully cloned from normal human colon mucosa. The vector pCMV5-myc-hTFF3 was reconstructed, and in 293-T cells transfected with the vector, hTFF3 expression was detected by Western blotting.
