Cloning and expression of mouse peroxiredoxin I in IEC-6 Cells.
OBJECTIVE
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.


CONCLUSIONS
Mouse peroxiredoxin I can be successfully expressed in IEC-6 cells.
Zhang
Bo
B
Institute of Combined Injury of PLA, State Key Laboratory of Trauma, Third Military Medical University, Gaotanyan Street 30, Chongqing 400038, China.
Su
Yong-Ping
YP
Wang
Tao
T
Wang
Feng-Chao
FC
Ai
Guo-Ping
GP
Xu
Hui
H
Wang
Jun-Ping
JP
Huang
Yue-Sheng
YS
Jiang
Jian-Xin
JX
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism*; D054464:Peroxiredoxins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D018076:DNA, Complementary; D010544:Peroxidases; D054464:Peroxiredoxins