2008•PubMedRequires access
[Construction of a recombinant eukaryotic vector of human intestinal trefoil factor and its expression in 293-T cells].
Ya-Pi Lu, Fei Zhou, Lin Wang, Bo Zhang, Jing Dong, Jianlin Ren
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Abstract
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.