Construction of a recombinant eukaryotic vector of human intestinal trefoil factor and its expression in 293-T cells.
- Author:
Ya-pi LU
1
;
Fei ZHOU
;
Lin WANG
;
Bo ZHANG
;
Jing DONG
;
Jian-lin REN
Author Information
- Publication Type:Journal Article
- MeSH: Blotting, Western; Cell Line; Eukaryotic Cells; metabolism; Genetic Vectors; genetics; Humans; Intestinal Mucosa; metabolism; Peptides; genetics; metabolism; Recombinant Proteins; biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; Transfection; methods; Trefoil Factor-2; Trefoil Factor-3
- From: Journal of Southern Medical University 2008;28(9):1630-1633
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone human intestinal trefoil factor (hITF/hTFF3) gene into an eukaryotic expression vector for its expression in eukaryotic cells.
METHODSThe 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 CONCLUSIONhTFF3 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.
