[Construction of a recombinant eukaryotic vector of human intestinal trefoil factor and its expression in 293-T cells].
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.


CONCLUSIONS
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.
Lu
Ya-pi
YP
Department of Gastroenterology, Zhongshan Hospital, Xiamen University, Xiamen, China. yapi@xmzsh.com
Zhou
Fei
F
Wang
Lin
L
Zhang
Bo
B
Dong
Jing
J
Ren
Jian-lin
JL
D015153:Blotting, Western; D002460:Cell Line; D005057:Eukaryotic Cells / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods; D000071164:Trefoil Factor-2; D000071165:Trefoil Factor-3
D010455:Peptides; D011994:Recombinant Proteins; C082495:TFF3 protein, human; D000071164:Trefoil Factor-2; D000071165:Trefoil Factor-3