Cloning and expression of mouse peroxiredoxin I in IEC-6 Cells.
World J Gastroenterol
; 10(14): 2109-12, 2004 Jul 15.
Article
in En
| MEDLINE
| ID: mdl-15237445
ABSTRACT
AIM:
To clone and express mouse peroxiredoxin I in IEC-6 cells.METHODS:
Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T-vector and sequenced, pSG5 was used to transiently express peroxiredoxin I in IEC-6 by liposome-mediated transfection, and the expression of peroxiredoxin I was evaluated by RT-PCR and Western blot.RESULTS:
A DNA fragment about 750 bp was amplified from total RNAs of IEC-6 cells using specific primers of peroxiredoxin I. The sequencing confirmed the coding region was successfully cloned into T-vector, which was completely coincident with the sequence in GeneBank. After the EcoRI-BamHI fragment of T-vector containing peroxiredoxin I was inserted into pSG5, the recombinant plasmid was transferred to IEC-6 cells. RT-PCR assay showed that a DNA fragment of 930 bp could be amplified, which indicated the transcription of pSG5-Prx. Western blot confirmed the expression of peroxiredoxin I in IEC-6 cells.CONCLUSION:
Mouse peroxiredoxin I can be successfully expressed in IEC-6 cells.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Peroxidases
/
Cloning, Molecular
Limits:
Animals
Language:
En
Journal:
World J Gastroenterol
Journal subject:
GASTROENTEROLOGIA
Year:
2004
Type:
Article
Affiliation country:
China