Construction of a recombinant eukaryotic vector of human intestinal trefoil factor and its expression in 293-T cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 1630-1633, 2008.
Article
in Zh
| WPRIM
| ID: wpr-340762
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To clone human intestinal trefoil factor (hITF/hTFF3) gene into an eukaryotic expression vector for its expression in eukaryotic cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS AND CONCLUSION</b>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.</p>
Full text:
1
Index:
WPRIM
Main subject:
Peptides
/
Recombinant Proteins
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Transfection
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Cell Line
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Blotting, Western
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Reverse Transcriptase Polymerase Chain Reaction
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Eukaryotic Cells
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Trefoil Factor-2
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Trefoil Factor-3
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Genetic Vectors
Limits:
Humans
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2008
Type:
Article