[Expression of human TFF3 in Escherichia coli].
OBJECTIVE
To construct recombinant human TFF3 prokaryotic expressing plasmid, express it in E. coli and identify the expressed protein.


METHODS
The cDNA for mature peptide of TFF3 was amplified by RT-PCR from RNA of human colon tissue and inserted into the MCS of the prokaryotic expressing plasmid pET32a (+). Then TFF3 was expressed as a fusion protein by IPTG induction. The recombinant protein was determined by SDS-PAGE and Western blot with a rabbit anti-TFF3 polyclonal antibody.


RESULTS
Sequencing result indicated that the obtained TFF3 fragment was inserted into plasmid pET32a (+) successfully and the sequence was correct. The expression level of the fusion protein was highest after 6h induction with 1 mmol/L IPTG. The result of Western blot demonstrated that the relative molecular mass of recombinant protein was about 24 x 10(3) and the protein had good antigenicity and specificity.


CONCLUSIONS
The expression plasmid pET32a-TFF3 was constructed and expressed successfully. This study will provide a substantial basis for further study of human TFF3.
Lu
Rui
R
Department of Parasitology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.
Wang
Xin
X
Chen
Jian-Ping
JP
Chen
Xian
X
Xu
Jia-Nan
JN
D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D000071165:Trefoil Factor-3
D010455:Peptides; D011993:Recombinant Fusion Proteins; C082495:TFF3 protein, human; D000071165:Trefoil Factor-3